Solubility data and modeling for sugar alcohols in ionic liquids

International Nuclear Information System (INIS)

Okuniewski, Marcin; Ramjugernath, Deresh; Naidoo, Paramespri; Domańska, Urszula

2014-01-01

Highlights: • Solubility of D-sorbitol and xylitol in six ILs. • The (liquid + liquid) phase equilibrium of (SA + IL) with UCST. • Interesting properties of [BMIM][TDI] IL. • The correlation with NRTL model. - Abstract: Ionic liquids (ILs) are novel media characterized by strong interactions with different organic substances which leads to a wide spectrum of applications involving extraction. Ionic liquids have been used as a solvent for sugar alcohols, sugars and hydrates. This work demonstrates the experimental and theoretical study of (liquid + liquid) phase equilibria for two sugar alcohols, D-sorbitol and xylitol in a few ILs based on different cations and anions (namely, 1-ethyl-1-methylpiperidinium bis(trifluoromethylsulfonyl)imide [EMPIP][NTf 2 ], 1-hexyl-1-methylpiperidinium bis(trifluoromethylsulfonyl)imide [HMPIP][NTf 2 ], N-hexylquinolinium bis(trifluoromethylsulfonyl)imide [HQuin][NTf 2 ], N-hexylisoquinolinium bis(trifluoromethylsulfonyl)imide [HiQuin][NTf 2 ], 1-butyl-1-methylimidazolium 4,5-dicyano-2-(trifluoromethyl)-imidazolide [BMIM][TDI] and 1-(cyanomethyl)-3-methylimidazolium 4,5-dicyano-2-(trifluoromethyl)-imidazolide [CCNMIM][TDI]). This study was conducted to assess the applicability of the studied ILs for dissolution of these biomass-related materials. (Liquid + liquid) phase equilibrium diagrams (LLE) in binary systems (sugar alcohol + ionic liquid) were measured using the dynamic technique. The influence of the chemical structure of both the ionic liquids and sugar alcohols were established and is discussed

Determination of soluble protein contents from RVNRL

International Nuclear Information System (INIS)

Wan Manshol Wan Zin; Nurulhuda Othman

1996-01-01

This project was carried out to determine the soluble protein contents on RVNRL film vulcanisates, with respect to the RVNRL storage time, gamma irradiation dose absorbed by the latex and the effect of different leaching time and leaching conditions. These three factors are important in the hope to determine the best possible mean of minimizing the soluble protein contents in products made from RVNRL. Within the nine months storage period employed in the study, the results show that, the longer the storage period the less the soluble protein extracted from the film samples. Gamma irradiation dose absorbed by the samples, between 5.3 kGy to 25.2 kGy seems to influence the soluble protein contents of the RVNRL films vulcanisates. The higher the dose the more was the soluble protein extracted from the film samples. At an absorbed dose of 5.3 kGy and 25.2 kGy, the soluble contents were 0. 198 mg/ml and 0.247 mg/ml respectively. At a fixed leaching temperature, the soluble proteins increases with leaching time and at a fixed leaching time, the soluble proteins increases with leaching temperature. ne highest extractable protein contents was determined at a leaching time of 10 minutes and leaching temperature of 90'C The protein analysis were done by using Modified Lowry Method

Soluble Sugars as the Carbohydrate Reserve for CAM in Pineapple Leaves 1

Science.gov (United States)

Carnal, Nancy Wieland; Black, Clanton C.

1989-01-01

Neutral ethanol-soluble sugar pools serve as carbohydrate reserves for Crassulacean acid metabolism (CAM) in pineapple (Ananas comosus (L.) Merr.) leaves. Levels of neutral soluble sugars and glucans fluctuated reciprocally with concentrations of malic acid. Hexose loss from neutral soluble-sugar pools was sufficient to account for malic acid accumulation with about 95% of the required hexose accounted for by turnover of fructose and glucose pools. Hexose loss from starch or starch plus lower molecular weight glucan pools was insufficient to account for nocturnal accumulation of malic acid. The apparent maximum catalytic capacity of pyrophosphate:6-phosphofructokinase (PPi-PFK) at 15°C was about 16 times higher than the mean maximum rate of glycolysis that occurred to support malic acid accumulation in pineapple leaves at night and 12 times higher than the mean maximum rate of hexose turnover from all carbohydrate pools. The apparent maximum catalytic capacity of ATP-PFK at 15°C was about 70% of the activity required to account for the mean maximal rate of hexose turnover from all carbohydrate pools if turnover were completely via glycolysis, and marginally sufficient to account for mean maximal rates of acidification. Therefore, at low night temperatures conducive to CAM and under subsaturating substrate concentrations, PPi-PFK activity, but not ATP-PFK activity, would be sufficient to support the rate of glycolytic carbohydrate processing required for acid accumulation. These data for pineapple establish that there are at least two types of CAM plants with respect to the nature of the carbohydrate reserve utilized to support nighttime CO2 accumulation. The data further indicate that the glycolytic carbohydrate processing that supports acidification proceeds in different subcellular compartments in plants utilizing different carbohydrate reserves. PMID:16666775

Seasonal patterns of reserve and soluble carbohydrates in mature sugar maple (Acer saccharum)

Science.gov (United States)

B.L. Wong; K.L. Baggett; A.H. Rye

2003-01-01

Sugar maple (Acer saccharum Marsh.) trees exhibit seasonal patterns of production, accumulation, and utilization of nonstructural carbohydrates that are closely correlated with phenological events and (or) physiological processes. The simultaneous seasonal patterns of both reserve and soluble carbohydrates in the leaves, twigs, branches, and trunks of healthy mature...

Countercurrent extraction of soluble sugars from almond hulls and assessment of the bioenergy potential.

Science.gov (United States)

Holtman, Kevin M; Offeman, Richard D; Franqui-Villanueva, Diana; Bayati, Andre K; Orts, William J

2015-03-11

Almond hulls contain considerable proportions (37% by dry weight) of water-soluble, fermentable sugars (sucrose, glucose, and fructose), which can be extracted for industrial purposes. The maximum optimal solids loading was determined to be 20% for sugar extraction, and the addition of 0.5% (w/v) pectinase aided in maintaining a sufficient free water volume for sugar recovery. A laboratory countercurrent extraction experiment utilizing a 1 h steep followed by three extraction (wash) stages produced a high-concentration (131 g/L fermentable sugar) syrup. Overall, sugar recovery efficiency was 88%. The inner stage washing efficiencies were compatible with solution equilibrium calculations, indicating that efficiency was high. The concentrated sugar syrup was fermented to ethanol at high efficiency (86% conversion), and ethanol concentrations in the broth were 7.4% (v/v). Thin stillage contained 233 g SCOD/L, which was converted to biomethane at an efficiency of 90% with a biomethane potential of 297 mL/g SCODdestroyed. Overall, results suggested that a minima of 49 gal (185 L) ethanol and 75 m(3) methane/t hulls (dry whole hull basis) are achievable.

Comparing side chain packing in soluble proteins, protein-protein interfaces, and transmembrane proteins.

Science.gov (United States)

Gaines, J C; Acebes, S; Virrueta, A; Butler, M; Regan, L; O'Hern, C S

2018-05-01

We compare side chain prediction and packing of core and non-core regions of soluble proteins, protein-protein interfaces, and transmembrane proteins. We first identified or created comparable databases of high-resolution crystal structures of these 3 protein classes. We show that the solvent-inaccessible cores of the 3 classes of proteins are equally densely packed. As a result, the side chains of core residues at protein-protein interfaces and in the membrane-exposed regions of transmembrane proteins can be predicted by the hard-sphere plus stereochemical constraint model with the same high prediction accuracies (>90%) as core residues in soluble proteins. We also find that for all 3 classes of proteins, as one moves away from the solvent-inaccessible core, the packing fraction decreases as the solvent accessibility increases. However, the side chain predictability remains high (80% within 30°) up to a relative solvent accessibility, rSASA≲0.3, for all 3 protein classes. Our results show that ≈40% of the interface regions in protein complexes are "core", that is, densely packed with side chain conformations that can be accurately predicted using the hard-sphere model. We propose packing fraction as a metric that can be used to distinguish real protein-protein interactions from designed, non-binding, decoys. Our results also show that cores of membrane proteins are the same as cores of soluble proteins. Thus, the computational methods we are developing for the analysis of the effect of hydrophobic core mutations in soluble proteins will be equally applicable to analyses of mutations in membrane proteins. © 2018 Wiley Periodicals, Inc.

Changes in physical, chemical and functional properties of whey protein isolate (WPI) and sugar beet pectin (SBP) conjugates formed by controlled dry-heating

Science.gov (United States)

A Maillard type reaction in the dry state was utilized to create conjugates between whey protein isolate (WPI) and sugar beet pectin (SBP) to achieve improved functional properties including solubility, colloidal stability and oil-in-water emulsion stability. To optimize the reaction conditions, mi...

Water deficit modifies the carbon isotopic composition of lipids, soluble sugars and leaves of Copaifera langsdorffii Desf. (Fabaceae

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Angelo Albano da Silva Bertholdi

2017-11-01

Full Text Available ABSTRACT Water deficit is most frequent in forest physiognomies subjected to climate change. As a consequence, several tree species alter tissue water potential, gas exchange and production of carbon compounds to overcome damage caused by water deficiency. The working hypothesis, that a reduction in gas exchange by plants experiencing water deficit will affect the composition of carbon compounds in soluble sugars, lipids and vegetative structures, was tested on Copaifera langsdorffii. Stomatal conductance, leaf water potential, and CO2 assimilation rate declined after a period of water deficit. After rehydration, leaf water potential and leaf gas exchange did not recover completely. Water deficit resulted in 13C enrichment in leaves, soluble sugars and root lipids. Furthermore, the amount of soluble sugars and root lipids decreased after water deficit. In rehydration, the carbon isotopic composition and amount of root lipids returned to levels similar to the control. Under water deficit, 13C-enriched in root lipids assists in the adjustment of cellular membrane turgidity and avoids damage to the process of water absorption by roots. These physiological adjustments permit a better understanding of the responses of Copaifera langsdorffi to water deficit.

Evaluation of the site specific protein glycation and antioxidant capacity of rare sugar-protein/peptide conjugates.

Science.gov (United States)

Sun, Yuanxia; Hayakawa, Shigeru; Ogawa, Masahiro; Izumori, Ken

2005-12-28

Protein-sugar conjugates generated in nonenzymatic glycation of alpha-lactalbumin (LA) with rare sugars [D-allose (All) and D-psicose (Psi)] and alimentary sugars as controls [D-glucose (Glc) and D-fructose (Fru)] were qualitatively determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Mass spectra revealed that the extent of glycation at lysine residues on LA with D-aldose molecules was very much higher than that of glycation with d-ketose molecules. To identify the specific site of glycation, the peptide mapping was established from protease V8 digestion, using a combination of computational cutting of proteins and MALDI-TOF-MS. As compared to peptide mapping, three and seven glycation sites were located in the primary structure of LA-ketose and LA-aldose conjugates, respectively. On the other hand, the antioxidant activities of protein-sugar conjugates and their peptic hydrolysates were investigated by 1,1-diphenyl-2-picrylhydrazyl radical scavenging method. The antioxidant activities of proteins/peptides glycated with rare sugars were significantly higher than those modified with the control sugars. The results indicated that the glycation degree and position were not markedly different between rare sugar and corresponding control sugar, but the antioxidant properties of protein and its hydrolysate were significantly enhanced by modifying with rare sugar.

The relationship between growth and soluble sugar concentration of Aloe vera plants grown under three levels of irradiance

Energy Technology Data Exchange (ETDEWEB)

Paez, A.; Gebre, G.M.; Tschaplinski, T.J. (Universidad del Zulia (Venezuela))

1994-06-01

The CAM plant Aloe vera was vegetatively propagated and grown under three irradiances: full sun, partial and deep shade (30% and 10% of ambient light, respectively) to determine the effect on growth, biomass allocation, and sugar concentration. After one year, the plants were harvested to determine final dry weight and the sugar concentration of the leaf mucilaginous gel. Plants grown under full sun produced twice the total dry weight of those grown under partial shade, with the difference equally partitioned between the shoot and root. Plants grown under full sun also produced thicker leaves, and more numerous and large auxiliary shoots. The dry weight of plants grown under deep shade was 8.6% that of plants grown under full sun, which was directly proportional to the irradiance received. Partial shade increased the number and length of leaves produced on the primary shoot, but the allocation of carbon to roots was the lowest of all treatments. Partial shade reduced the total sugar concentration of the leaf gel matrix to 34% that of plants under full sun, due to reductions in all sugars measured. Glucose was the most abundant soluble sugar, with its relative contribution to the total pool increasing under shade. In summary, the proportional effects of partial shading were greater on soluble sugar concentrations than on the total plant biomass produced.

Predicting the solubility of mixtures of sugars and their replacers using the Flory-Huggins theory

NARCIS (Netherlands)

Sman, van der R.G.M.

2017-01-01

In this paper we investigate whether the Flory-Huggins theory can describe the thermodynamics of solutions of simple carbohydrates, like sugars and polyols. In particular, we focus on the description of the solubility of the carbohydrates in water. This is investigated for both binary and ternary

Chemical constituents: water-soluble vitamins, free amino acids and sugar profile from Ganoderma adspersum.

Science.gov (United States)

Kıvrak, İbrahim

2015-01-01

Ganoderma adspersum presents a rigid fruiting body owing to chitin content and having a small quantity of water or moisture. The utility of bioactive constituent of the mushroom can only be available by extraction for human usage. In this study, carbohydrate, water-soluble vitamin compositions and amino acid contents were determined in G. adspersum mushroom. The composition in individual sugars was determined by HPLC-RID, mannitol (13.04 g/100 g) and trehalose (10.27 g/100 g) being the most abundant sugars. The examination of water-soluble vitamins and free amino acid composition was determined by UPLC-ESI-MS/MS. Essential amino acid constituted 67.79% of total amino acid, which is well worth the attention with regard to researchers and consumers. In addition, G. adspersum, which is also significantly rich in B group vitamins and vitamin C, can provide a wide range of notable applications in the pharmaceutics, cosmetics, food and dietary supplement industries. G. adspersum revealed its value for pharmacy and nutrition fields.

X-radiation effect on soluble proteins of gastric mucosa

International Nuclear Information System (INIS)

Sukhomlinov, B.F.; Chajka, Ya.P.; Fedorovich, A.N.

1979-01-01

Using the method of electrophoresis in agar gel soluble proteins of gastric mucosa of rats were separated into 11 fractions. Proteins posessing a proteolytic (pH 1.8) and lipase (pH 7.4) activity were localized within the second and third prealbumin fractions. Soluble proteins of gastric mucosa contain glyco- and lipoproteid complexes. Exposure of rats to 1000 R of X-rays induces quantitative redistribution within the electrophoretic spectrum of soluble proteins and a considerable disturbance of the proteolytic activity of total soluble proteins throughout the entire period of observation (from 10 min to 72h)

Characterization of soluble protein BCP 11/24 from bovine corneal epithelium, different from the principal soluble protein BCP 54

NARCIS (Netherlands)

Bakker, C.; Pasmans, S.; Verhagen, C.; van Haren, M.; van der Gaag, R.; Hoekzema, R.

1992-01-01

The water-soluble fraction of bovine corneal epithelium was analysed by polyacrylamide gel electrophoresis in the presence of SDS (SDS-PAGE). Next to the principal soluble protein BCP 54, which has recently been identified as a corneal aldehyde dehydrogenase (ALDH), another abundant protein was

Sugar regulation of SUGAR TRANSPORTER PROTEIN 1 (STP1) expression in Arabidopsis thaliana

Science.gov (United States)

Cordoba, Elizabeth; Aceves-Zamudio, Denise Lizeth; Hernández-Bernal, Alma Fabiola; Ramos-Vega, Maricela; León, Patricia

2015-01-01

Sugars regulate the expression of many genes at the transcriptional level. In Arabidopsis thaliana, sugars induce or repress the expression of >1800 genes, including the STP1 (SUGAR TRANSPORTER PROTEIN 1) gene, which encodes an H+/monosaccharide cotransporter. STP1 transcript levels decrease more rapidly after the addition of low concentrations of sugars than the levels of other repressed genes, such as DIN6 (DARK-INDUCED 6). We found that this regulation is exerted at the transcriptional level and is initiated by phosphorylatable sugars. Interestingly, the sugar signal that modulates STP1 expression is transmitted through a HEXOKINASE 1-independent signalling pathway. Finally, analysis of the STP1 5′ regulatory region allowed us to delimit a region of 309bp that contains the cis elements implicated in the glucose regulation of STP1 expression. Putative cis-acting elements involved in this response were identified. PMID:25281700

Relationship between Molecular Structure Characteristics of Feed Proteins and Protein Digestibility and Solubility

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Mingmei Bai

2016-08-01

Full Text Available The nutritional value of feed proteins and their utilization by livestock are related not only to the chemical composition but also to the structure of feed proteins, but few studies thus far have investigated the relationship between the structure of feed proteins and their solubility as well as digestibility in monogastric animals. To address this question we analyzed soybean meal, fish meal, corn distiller’s dried grains with solubles, corn gluten meal, and feather meal by Fourier transform infrared (FTIR spectroscopy to determine the protein molecular spectral band characteristics for amides I and II as well as α-helices and β-sheets and their ratios. Protein solubility and in vitro digestibility were measured with the Kjeldahl method using 0.2% KOH solution and the pepsin-pancreatin two-step enzymatic method, respectively. We found that all measured spectral band intensities (height and area of feed proteins were correlated with their the in vitro digestibility and solubility (p≤0.003; moreover, the relatively quantitative amounts of α-helices, random coils, and α-helix to β-sheet ratio in protein secondary structures were positively correlated with protein in vitro digestibility and solubility (p≤0.004. On the other hand, the percentage of β-sheet structures was negatively correlated with protein in vitro digestibility (p<0.001 and solubility (p = 0.002. These results demonstrate that the molecular structure characteristics of feed proteins are closely related to their in vitro digestibility at 28 h and solubility. Furthermore, the α-helix-to-β-sheet ratio can be used to predict the nutritional value of feed proteins.

Model to predict inhomogeneous protein-sugar distribution in powders prepared by spray drying

NARCIS (Netherlands)

Grasmeijer, Niels; Frijlink, Henderik W.; Hinrichs, Wouter L. J.

2016-01-01

A protein can be stabilized by spray drying an aqueous solution of the protein and a sugar, thereby incorporating the protein into a glassy sugar matrix. For optimal stability, the protein should be homogeneously distributed inside the sugar matrix. The aim of this study was to develop a model that

Lysine Rich Proteins in the Salt-Soluble Protein Fraction of Barley

DEFF Research Database (Denmark)

Ingversen, J.; Køie, B.

1973-01-01

Fractionation of the protein complex from Emir barley showed that the salt-soluble fraction accounts for 44% of the total lysine content but only for 2.......Fractionation of the protein complex from Emir barley showed that the salt-soluble fraction accounts for 44% of the total lysine content but only for 2....

Effects of putrescine, kinetin and IAA on protein synthesis in 'Phaseolus vulgaris' coleoptiles

Energy Technology Data Exchange (ETDEWEB)

Crocomo, O J; Lee, T S.G. [Centro de Energia Nuclear na Agricultura, Piracicaba (Brazil)

1975-01-01

Incubation of etiolated 'Phaseolus vulgaris' coleoptiles shows a converse flux between soluble protein and reducing sugar. The rate of incorporation of radioactive arginine into protein was higher than that of radioactive leucine. Radioactive arginine incorporation into protein was linear up to 120 min and then started to decline. The rate of incorporation of radioactive leucine was increased by preincubation of the tissue in the incubation medium. Roots were found to contain more soluble protein and much less reducing sugar than the coleoptile. The optimum pH value for protein synthesis in coleoptile sections was found to be 6 for control tissues and 4 for those treated with 10-/sup 3/M IAA. This high concentration of IAA was also found to inhibit soluble protein synthesis, the incorporation rate of radioactive arginine and leucine into protein fraction, the secretion of hydrogen ion into the incubation medium and elongation of the bean segment. Kinetin at 2x10/sup -4/M and putrescine at 5mM both decreased the rate of /sup 14/C-arginine incorporation into soluble protein, but for /sup 14/C-leucine, this rate of incorporation was found to be increased after 90 min incubation with a preincubation of 30 min. In general, the change pattern of the soluble protein content, the reducing sugar level and the incorporation rate of radioactive arginine and leucine into protein in the kinetin and putrescine treated tissues were about the same although tissues that incubated with kinetin always contain more soluble protein and less reducing sugar than that of incubated with putrescine.

Generation of Soluble Advanced Glycation End Products Receptor (sRAGE)-Binding Ligands during Extensive Heat Treatment of Whey Protein/Lactose Mixtures Is Dependent on Glycation and Aggregation

NARCIS (Netherlands)

Liu, Fahui; Teodorowicz, Gosia; Wichers, Harry J.; Boekel, van Tiny; Hettinga, Kasper A.

2016-01-01

Heating of protein- and sugar-containing materials is considered the primary factor affecting the formation of advanced glycation end products (AGEs). This study aimed to investigate the influence of heating conditions, digestion, and aggregation on the binding capacity of AGEs to the soluble AGE

Two novel heat-soluble protein families abundantly expressed in an anhydrobiotic tardigrade.

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Ayami Yamaguchi

Full Text Available Tardigrades are able to tolerate almost complete dehydration by reversibly switching to an ametabolic state. This ability is called anhydrobiosis. In the anhydrobiotic state, tardigrades can withstand various extreme environments including space, but their molecular basis remains largely unknown. Late embryogenesis abundant (LEA proteins are heat-soluble proteins and can prevent protein-aggregation in dehydrated conditions in other anhydrobiotic organisms, but their relevance to tardigrade anhydrobiosis is not clarified. In this study, we focused on the heat-soluble property characteristic of LEA proteins and conducted heat-soluble proteomics using an anhydrobiotic tardigrade. Our heat-soluble proteomics identified five abundant heat-soluble proteins. All of them showed no sequence similarity with LEA proteins and formed two novel protein families with distinct subcellular localizations. We named them Cytoplasmic Abundant Heat Soluble (CAHS and Secretory Abundant Heat Soluble (SAHS protein families, according to their localization. Both protein families were conserved among tardigrades, but not found in other phyla. Although CAHS protein was intrinsically unstructured and SAHS protein was rich in β-structure in the hydrated condition, proteins in both families changed their conformation to an α-helical structure in water-deficient conditions as LEA proteins do. Two conserved repeats of 19-mer motifs in CAHS proteins were capable to form amphiphilic stripes in α-helices, suggesting their roles as molecular shield in water-deficient condition, though charge distribution pattern in α-helices were different between CAHS and LEA proteins. Tardigrades might have evolved novel protein families with a heat-soluble property and this study revealed a novel repertoire of major heat-soluble proteins in these anhydrobiotic animals.

Relationship between Molecular Structure Characteristics of Feed Proteins and Protein In vitro Digestibility and Solubility.

Science.gov (United States)

Bai, Mingmei; Qin, Guixin; Sun, Zewei; Long, Guohui

2016-08-01

The nutritional value of feed proteins and their utilization by livestock are related not only to the chemical composition but also to the structure of feed proteins, but few studies thus far have investigated the relationship between the structure of feed proteins and their solubility as well as digestibility in monogastric animals. To address this question we analyzed soybean meal, fish meal, corn distiller's dried grains with solubles, corn gluten meal, and feather meal by Fourier transform infrared (FTIR) spectroscopy to determine the protein molecular spectral band characteristics for amides I and II as well as α-helices and β-sheets and their ratios. Protein solubility and in vitro digestibility were measured with the Kjeldahl method using 0.2% KOH solution and the pepsin-pancreatin two-step enzymatic method, respectively. We found that all measured spectral band intensities (height and area) of feed proteins were correlated with their the in vitro digestibility and solubility (p≤0.003); moreover, the relatively quantitative amounts of α-helices, random coils, and α-helix to β-sheet ratio in protein secondary structures were positively correlated with protein in vitro digestibility and solubility (p≤0.004). On the other hand, the percentage of β-sheet structures was negatively correlated with protein in vitro digestibility (pdigestibility at 28 h and solubility. Furthermore, the α-helix-to-β-sheet ratio can be used to predict the nutritional value of feed proteins.

Effects of bagging on sugar metabolism and the activity of sugar ...

African Journals Online (AJOL)

To investigate the effects of bagging on sugar metabolism and the activity of sugar metabolism related enzymes in Qingzhong loquat fruit development, the contents of sucrose, glucose and soluble solids as well as the activities of sugar metabolism related enzymes were evaluated. The content of sucrose, glucose and ...

Facilitating protein solubility by use of peptide extensions

Science.gov (United States)

Freimuth, Paul I; Zhang, Yian-Biao; Howitt, Jason

2013-09-17

Expression vectors for expression of a protein or polypeptide of interest as a fusion product composed of the protein or polypeptide of interest fused at one terminus to a solubility enhancing peptide extension are provided. Sequences encoding the peptide extensions are provided. The invention further comprises antibodies which bind specifically to one or more of the solubility enhancing peptide extensions.

Impact of the nutrients N and K and soluble sugars on Diabrotica speciosa (Germar) (Coleoptera, Chrysomelidae) and Agrotis ipsilon (Hufnagel) (Lepidoptera, Noctuidae) populations in potato crops, Solanum tuberosum L. (Solanaceae)

International Nuclear Information System (INIS)

Azeredo, Edson Henrique de; Lima, Eduardo; Cassino, Paulo Cesar Rodrigues

2004-01-01

Impact of the nutrients N and K and soluble sugars on Diabrotica speciosa (Germar) (Coleoptera, Chrysomelidae) and Agrotis ipsilon (Huefnagel) (Lepidoptera, Noctuidae) populations in potato crops, Solanum tuberosum L. (Solanaceae). The occurrence of Diabrotica speciosa (Germar, 1824) and Agrotis ipsilon (Huefnagel, 1767) on the potato cultivars Achat and Monalisa, influenced by nitrogen and potassium dosage, and minimum quantity of soluble sugars, was studied. The following parameters were evaluated: concentration of mineral nutrient and sugar in green leaf, senescent leaf, leaf in abscission, stem, tubercle and total plant using extracts of infusion in ethanol 80%. The largest infestation of D. speciosa larvae was on Monalisa cultivar at 150 kg.ha -1 of N + K with 27.03% at P -1 , in the absence of potassium. On the other hand, high dosage of K reduced the damages by A. ipsilon on Monalisa cultivar. However, it did not influence the storage of soluble sugar. The results indicated that in Achat cultivar the accumulated soluble sugar was reduced, probably sensitized by elevation of potassic fertilization dosing, differing from Monalisa cultivar, in which the influence was by nitrogen dosing. (author)

EFFECT OF HEAT TREATMENT ON SOYBEAN PROTEIN SOLUBILITY

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RODICA CĂPRIŢĂ

2007-05-01

Full Text Available The use of soybean products in animal feeds is limited due to the presence of antinutritional factors (ANF. Proper heat processing is required to destroy ANF naturally present in raw soybeans and to remove solvent remaining from the oil extraction process. Over and under toasting of soybean causes lower nutritional value. Excessive heat treatment causes Maillard reaction which affects the availability of lysine in particular and produces changes to the chemical structure of proteins resulting in a decrease of the nutritive value. The objective of this study was to evaluate the effect of heating time on the protein solubility. The investigation of the heating time on protein solubility in soybean meal (SBM revealed a negative correlation (r = -0.9596. Since the urease index is suitable only for detecting under processed SBM, the protein solubility is an important index for monitoring SBM quality.

Self assembling nanocomposites for protein delivery: supramolecular interactions of soluble polymers with protein drugs.

Science.gov (United States)

Salmaso, Stefano; Caliceti, Paolo

2013-01-02

Translation of therapeutic proteins to pharmaceutical products is often encumbered by their inadequate physicochemical and biopharmaceutical properties, namely low stability and poor bioavailability. Over the last decades, several academic and industrial research programs have been focused on development of biocompatible polymers to produce appropriate formulations that provide for enhanced therapeutic performance. According to their physicochemical properties, polymers have been exploited to obtain a variety of formulations including biodegradable microparticles, 3-dimensional hydrogels, bioconjugates and soluble nanocomposites. Several soluble polymers bearing charges or hydrophobic moieties along the macromolecular backbone have been found to physically associate with proteins to form soluble nanocomplexes. Physical complexation is deemed a valuable alternative tool to the chemical bioconjugation. Soluble protein/polymer nanocomplexes formed by physical specific or unspecific interactions have been found in fact to possess peculiar physicochemical, and biopharmaceutical properties. Accordingly, soluble polymeric systems have been developed to increase the protein stability, enhance the bioavailability, promote the absorption across the biological barriers, and prolong the protein residence in the bloodstream. Furthermore, a few polymers have been found to favour the protein internalisation into cells or boost their immunogenic potential by acting as immunoadjuvant in vaccination protocols. Copyright © 2011 Elsevier B.V. All rights reserved.

Membrane Proteins Are Dramatically Less Conserved than Water-Soluble Proteins across the Tree of Life.

Science.gov (United States)

Sojo, Victor; Dessimoz, Christophe; Pomiankowski, Andrew; Lane, Nick

2016-11-01

Membrane proteins are crucial in transport, signaling, bioenergetics, catalysis, and as drug targets. Here, we show that membrane proteins have dramatically fewer detectable orthologs than water-soluble proteins, less than half in most species analyzed. This sparse distribution could reflect rapid divergence or gene loss. We find that both mechanisms operate. First, membrane proteins evolve faster than water-soluble proteins, particularly in their exterior-facing portions. Second, we demonstrate that predicted ancestral membrane proteins are preferentially lost compared with water-soluble proteins in closely related species of archaea and bacteria. These patterns are consistent across the whole tree of life, and in each of the three domains of archaea, bacteria, and eukaryotes. Our findings point to a fundamental evolutionary principle: membrane proteins evolve faster due to stronger adaptive selection in changing environments, whereas cytosolic proteins are under more stringent purifying selection in the homeostatic interior of the cell. This effect should be strongest in prokaryotes, weaker in unicellular eukaryotes (with intracellular membranes), and weakest in multicellular eukaryotes (with extracellular homeostasis). We demonstrate that this is indeed the case. Similarly, we show that extracellular water-soluble proteins exhibit an even stronger pattern of low homology than membrane proteins. These striking differences in conservation of membrane proteins versus water-soluble proteins have important implications for evolution and medicine. © The Author 2016. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

Accumulation of soluble sugars in peel at high temperature leads to stay-green ripe banana fruit.

Science.gov (United States)

Yang, Xiaotang; Pang, Xuequn; Xu, Lanying; Fang, Ruiqiu; Huang, Xuemei; Guan, Peijian; Lu, Wangjin; Zhang, Zhaoqi

2009-01-01

Bananas (Musa acuminata, AAA group) fail to develop a yellow peel and stay green when ripening at temperatures >24 degrees C. The identification of the mechanisms leading to the development of stay-green ripe bananas has practical value and is helpful in revealing pathways involved in the regulation of chlorophyll (Chl) degradation. In the present study, the Chl degradation pathway was characterized and the progress of ripening and senescence was assessed in banana peel at 30 degrees C versus 20 degrees C, by monitoring relevant gene expression and ripening and senescence parameters. A marked reduction in the expression levels of the genes for Chl b reductase, SGR (Stay-green protein), and pheophorbide a oxygenase was detected for the fruit ripening at 30 degrees C, when compared with fruit at 20 degrees C, indicating that Chl degradation was repressed at 30 degrees C at various steps along the Chl catabolic pathway. The repressed Chl degradation was not due to delayed ripening and senescence, since the fruit at 30 degrees C displayed faster onset of various ripening and senescence symptoms, suggesting that the stay-green ripe bananas are of similar phenotype to type C stay-green mutants. Faster accumulation of high levels of fructose and glucose in the peel at 30 degrees C prompted investigation of the roles of soluble sugars in Chl degradation. In vitro incubation of detached pieces of banana peel showed that the pieces of peel stayed green when incubated with 150 mM glucose or fructose, but turned completely yellow in the absence of sugars or with 150 mM mannitol, at either 20 degrees C or 30 degrees C. The results suggest that accumulation of sugars in the peel induced by a temperature of 30 degrees C may be a major factor regulating Chl degradation independently of fruit senescence.

A Soluble, Folded Protein without Charged Amino Acid Residues

DEFF Research Database (Denmark)

Højgaard, Casper; Kofoed, Christian; Espersen, Roall

2016-01-01

Charges are considered an integral part of protein structure and function, enhancing solubility and providing specificity in molecular interactions. We wished to investigate whether charged amino acids are indeed required for protein biogenesis and whether a protein completely free of titratable...... side chains can maintain solubility, stability, and function. As a model, we used a cellulose-binding domain from Cellulomonas fimi, which, among proteins of more than 100 amino acids, presently is the least charged in the Protein Data Bank, with a total of only four titratable residues. We find...

Co-extraction of soluble and insoluble sugars from energy sorghum based on a hydrothermal hydrolysis process.

Science.gov (United States)

Yu, Qiang; Tan, Xuesong; Zhuang, Xinshu; Wang, Qiong; Wang, Wen; Qi, Wei; Zhou, Guixiong; Luo, Yu; Yuan, Zhenhong

2016-12-01

A process for co-extraction of soluble and insoluble sugars from energy sorghum (ES) was developed based on hydrothermal hydrolysis (HH). Two series of ES were investigated: one (N) with a high biomass yield displayed a higher recalcitrance to sugar release, whereas the second (T) series was characterized by high sugar extraction. The highest total xylose recoveries of 87.2% and 98.7% were obtained for N-11 and T-106 under hydrolysis conditions of 180°C for 50min and 180°C for 30min, respectively. Moreover, the T series displayed higher enzymatic digestibility (ED) than the N series. The high degree of branching (arabinose/xylose ratio) and acetyl groups in the hemicellulose chains of T-106 would be expected to accelerate sugar release during the HH process. In addition, negative correlations between ED and the lignin content, crystallinity index (CrI) and syringyl/guaiacyl (S/G) lignin ratio were observed. Furthermore, finding ways to overcome the thickness of the cell wall and heterogeneity of its chemical composition distribution would make cellulose more accessible to the enzyme. Copyright © 2016 Elsevier Ltd. All rights reserved.

Production of soluble mammalian proteins in Escherichia coli: identification of protein features that correlate with successful expression

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Perera Rajika L

2004-12-01

Full Text Available Abstract Background In the search for generic expression strategies for mammalian protein families several bacterial expression vectors were examined for their ability to promote high yields of soluble protein. Proteins studied included cell surface receptors (Ephrins and Eph receptors, CD44, kinases (EGFR-cytoplasmic domain, CDK2 and 4, proteases (MMP1, CASP2, signal transduction proteins (GRB2, RAF1, HRAS and transcription factors (GATA2, Fli1, Trp53, Mdm2, JUN, FOS, MAD, MAX. Over 400 experiments were performed where expression of 30 full-length proteins and protein domains were evaluated with 6 different N-terminal and 8 C-terminal fusion partners. Expression of an additional set of 95 mammalian proteins was also performed to test the conclusions of this study. Results Several protein features correlated with soluble protein expression yield including molecular weight and the number of contiguous hydrophobic residues and low complexity regions. There was no relationship between successful expression and protein pI, grand average of hydropathicity (GRAVY, or sub-cellular location. Only small globular cytoplasmic proteins with an average molecular weight of 23 kDa did not require a solubility enhancing tag for high level soluble expression. Thioredoxin (Trx and maltose binding protein (MBP were the best N-terminal protein fusions to promote soluble expression, but MBP was most effective as a C-terminal fusion. 63 of 95 mammalian proteins expressed at soluble levels of greater than 1 mg/l as N-terminal H10-MBP fusions and those that failed possessed, on average, a higher molecular weight and greater number of contiguous hydrophobic amino acids and low complexity regions. Conclusions By analysis of the protein features identified here, this study will help predict which mammalian proteins and domains can be successfully expressed in E. coli as soluble product and also which are best targeted for a eukaryotic expression system. In some cases

Protein solubility and folding enhancement by interaction with RNA.

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Seong Il Choi

Full Text Available While basic mechanisms of several major molecular chaperones are well understood, this machinery has been known to be involved in folding of only limited number of proteins inside the cells. Here, we report a chaperone type of protein folding facilitated by interaction with RNA. When an RNA-binding module is placed at the N-terminus of aggregation-prone target proteins, this module, upon binding with RNA, further promotes the solubility of passenger proteins, potentially leading to enhancement of proper protein folding. Studies on in vitro refolding in the presence of RNA, coexpression of RNA molecules in vivo and the mutants with impaired RNA binding ability suggests that RNA can exert chaperoning effect on their bound proteins. The results suggest that RNA binding could affect the overall kinetic network of protein folding pathway in favor of productive folding over off-pathway aggregation. In addition, the RNA binding-mediated solubility enhancement is extremely robust for increasing soluble yield of passenger proteins and could be usefully implemented for high-throughput protein expression for functional and structural genomic research initiatives. The RNA-mediated chaperone type presented here would give new insights into de novo folding in vivo.

Optimization of translation profiles enhances protein expression and solubility.

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Anne-Katrin Hess

Full Text Available mRNA is translated with a non-uniform speed that actively coordinates co-translational folding of protein domains. Using structure-based homology we identified the structural domains in epoxide hydrolases (EHs and introduced slow-translating codons to delineate the translation of single domains. These changes in translation speed dramatically improved the solubility of two EHs of metagenomic origin in Escherichia coli. Conversely, the importance of transient attenuation for the folding, and consequently solubility, of EH was evidenced with a member of the EH family from Agrobacterium radiobacter, which partitions in the soluble fraction when expressed in E. coli. Synonymous substitutions of codons shaping the slow-transiting regions to fast-translating codons render this protein insoluble. Furthermore, we show that low protein yield can be enhanced by decreasing the free folding energy of the initial 5'-coding region, which can disrupt mRNA secondary structure and enhance ribosomal loading. This study provides direct experimental evidence that mRNA is not a mere messenger for translation of codons into amino acids but bears an additional layer of information for folding, solubility and expression level of the encoded protein. Furthermore, it provides a general frame on how to modulate and fine-tune gene expression of a target protein.

Atmospheric carbon dioxide changes photochemical activity, soluble sugars and volatile levels in broccoli (Brassica oleracea var. italica).

Science.gov (United States)

Krumbein, Angelika; Kläring, Hans-Peter; Schonhof, Ilona; Schreiner, Monika

2010-03-24

Atmospheric carbon dioxide (CO(2)) concentration is an environmental factor currently undergoing dramatic changes. The objective of the present study was to determine the effect of doubling the ambient CO(2) concentration on plant photochemistry as measured by photochemical quenching coefficient (qP), soluble sugars and volatiles in broccoli. Elevated CO(2) concentration increased qP values in leaves by up to 100% and 89% in heads, while glucose and sucrose in leaves increased by about 60%. Furthermore, in broccoli heads elevated CO(2) concentration induced approximately a 2-fold increase in concentrations of three fatty acid-derived C(7) aldehydes ((E)-2-heptenal, (E,Z)-2,4-heptadienal, (E,E)-2,4-heptadienal), two fatty acid-derived C(5) alcohols (1-penten-3-ol, (Z)-2-pentenol), and two amino acid-derived nitriles (phenyl propanenitrile, 3-methyl butanenitrile). In contrast, concentrations of the sulfur-containing compound 2-ethylthiophene and C(6) alcohol (E)-2-hexenol decreased. Finally, elevated CO(2) concentration increased soluble sugar concentrations due to enhanced photochemical activity in leaves and heads, which may account for the increased synthesis of volatiles.

Protein Solubility as Quality Index for Processed Soybean

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Rodica Căpriţă

2010-05-01

Full Text Available Protein quality of soybean meal (SBM is linked to both the reduction of antinutritional factors (ANFs, and the optimization of protein digestibility. Both insufficient- and over-heating result in poor quality SBM. Inadequate heating fails to completely destroy the ANFs, which may have a detrimental impact on animal performance, while excessive heating reduces the availability of lysine via the Maillard reaction and possibly, to a lesser extent, of other amino acids. The objective of our study was to compare some biochemical laboratory procedures for assessing quality of SBM: urease index (UI, protein dispersibility index (PDI, KOH protein solubility (PS, and nitrogen solubility index (NSI. The experimental data reveal that UI is not useful to determine excessive heat treatment since additional heating has no effect on the urease index. KOH protein solubility remains high, during initial heat treatment. In marked contrast, the PDI and NSI decreased incrementally from 78% to 20% and from 97% to 60%, respectively, when heating 0 to 30 minutes. Combing the PDI test with the urease test could be useful to monitor soybean quality. SBM containing low UI (0.3 or below and high PDI (40 to 45% may indicate that the sample is definitely high quality because it has been adequately heat processed, but not overprocessed.

PON-Sol: prediction of effects of amino acid substitutions on protein solubility.

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Yang, Yang; Niroula, Abhishek; Shen, Bairong; Vihinen, Mauno

2016-07-01

Solubility is one of the fundamental protein properties. It is of great interest because of its relevance to protein expression. Reduced solubility and protein aggregation are also associated with many diseases. We collected from literature the largest experimentally verified solubility affecting amino acid substitution (AAS) dataset and used it to train a predictor called PON-Sol. The predictor can distinguish both solubility decreasing and increasing variants from those not affecting solubility. PON-Sol has normalized correct prediction ratio of 0.491 on cross-validation and 0.432 for independent test set. The performance of the method was compared both to solubility and aggregation predictors and found to be superior. PON-Sol can be used for the prediction of effects of disease-related substitutions, effects on heterologous recombinant protein expression and enhanced crystallizability. One application is to investigate effects of all possible AASs in a protein to aid protein engineering. PON-Sol is freely available at http://structure.bmc.lu.se/PON-Sol The training and test data are available at http://structure.bmc.lu.se/VariBench/ponsol.php mauno.vihinen@med.lu.se Supplementary data are available at Bioinformatics online. © The Author 2016. Published by Oxford University Press. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.

A Soluble, Folded Protein without Charged Amino Acid Residues

DEFF Research Database (Denmark)

Højgaard, Casper; Kofoed, Christian; Espersen, Roall

2016-01-01

side chains can maintain solubility, stability, and function. As a model, we used a cellulose-binding domain from Cellulomonas fimi, which, among proteins of more than 100 amino acids, presently is the least charged in the Protein Data Bank, with a total of only four titratable residues. We find......Charges are considered an integral part of protein structure and function, enhancing solubility and providing specificity in molecular interactions. We wished to investigate whether charged amino acids are indeed required for protein biogenesis and whether a protein completely free of titratable...... that the protein shows a surprising resilience toward extremes of pH, demonstrating stability and function (cellulose binding) in the pH range from 2 to 11. To ask whether the four charged residues present were required for these properties of this protein, we altered them to nontitratable ones. Remarkably...

Efficient protein production method for NMR using soluble protein tags with cold shock expression vector

International Nuclear Information System (INIS)

Hayashi, Kokoro; Kojima, Chojiro

2010-01-01

The E. coli protein expression system is one of the most useful methods employed for NMR sample preparation. However, the production of some recombinant proteins in E. coli is often hampered by difficulties such as low expression level and low solubility. To address these problems, a modified cold-shock expression system containing a glutathione S-transferase (GST) tag, the pCold-GST system, was investigated. The pCold-GST system successfully expressed 9 out of 10 proteins that otherwise could not be expressed using a conventional E. coli expression system. Here, we applied the pCold-GST system to 84 proteins and 78 proteins were successfully expressed in the soluble fraction. Three other cold-shock expression systems containing a maltose binding protein tag (pCold-MBP), protein G B1 domain tag (pCold-GB1) or thioredoxin tag (pCold-Trx) were also developed to improve the yield. Additionally, we show that a C-terminal proline tag, which is invisible in 1 H- 15 N HSQC spectra, inhibits protein degradation and increases the final yield of unstable proteins. The purified proteins were amenable to NMR analyses. These data suggest that pCold expression systems combined with soluble protein tags can be utilized to improve the expression and purification of various proteins for NMR analysis.

Limitations of polyethylene glycol-induced precipitation as predictive tool for protein solubility during formulation development.

Science.gov (United States)

Hofmann, Melanie; Winzer, Matthias; Weber, Christian; Gieseler, Henning

2018-05-01

Polyethylene glycol (PEG)-induced protein precipitation is often used to extrapolate apparent protein solubility at specific formulation compositions. The procedure was used for several fields of application such as protein crystal growth but also protein formulation development. Nevertheless, most studies focused on applicability in protein crystal growth. In contrast, this study focuses on applicability of PEG-induced precipitation during high-concentration protein formulation development. In this study, solubility of three different model proteins was investigated over a broad range of pH. Solubility values predicted by PEG-induced precipitation were compared to real solubility behaviour determined by either turbidity or content measurements. Predicted solubility by PEG-induced precipitation was confirmed for an Fc fusion protein and a monoclonal antibody. In contrast, PEG-induced precipitation failed to predict solubility of a single-domain antibody construct. Applicability of PEG-induced precipitation as indicator of protein solubility during formulation development was found to be not valid for one of three model molecules. Under certain conditions, PEG-induced protein precipitation is not valid for prediction of real protein solubility behaviour. The procedure should be used carefully as tool for formulation development, and the results obtained should be validated by additional investigations. © 2017 Royal Pharmaceutical Society.

Relationships between soluble sugar concentrations in roots and ecosystem stress for first-year sugar maple seedlings

Energy Technology Data Exchange (ETDEWEB)

McLaughlin, J.W.; Reed, D.D.; Jurgensen, M.F.; Mroz, G.D.; Bagley, S.T. [Michigan Technological University, Houghton, MI (United States). School of Forestry and Wood Products

1996-03-01

Accumulation of reducing sugars (i.e. glucose and fructose) in plant roots has been consistently correlated with forest dieback and decline and, therefore, has potential as a biological indicator of ecosystem stress. In this study, the relationships between acidic deposition and `natural` (temperature, mycorrhizae, and nutrition) factors with first-year sugar maple seedling root sugar concentrations and growth were assessed in two sugar maple dominated forests in Michigan. Seedlings at the southern site (Wellston) had greater root growth, phosphorus, total sugar, and sucrose concentrations in roots, but lower reducing sugar concentration in roots. In addition, percent root length colonized by vesicular-arbuscular mycorrhizal fungi was less than that found for seedlings growing at the northern site (Alberta). Throughfall deposition of nitrate, sulfate, and hydrogen ions was not significantly correlated with seedling total or reducing sugar concentration. Total sugar concentration in seedling roots was positively correlated with air and soil temperatures at the southern site, but not at the northern site. Seedling tissue phosphorus concentration was correlated with total sugars at both sites, with sucrose at the southern site, and reducing sugars at the northern site. Mycorrhizal colonization rates at the Alberta site were positively correlated with reducing sugar concentration in seedling roots and negatively correlated with sucrose concentration. The results suggest that differences in seedling root sugar concentrations in these two forests are related to seedling root growth and are most likely due to ecological variables, such as available soil phosphorus, temperature, and growing season length through some complex interaction with mycorrhizae rather than acidic deposition stress. 56 refs., 3 figs.

The Influence of Tissue Lyophilization and Gamma Irradiation on the Solubility of Proteins

International Nuclear Information System (INIS)

Komender, J.; Jendyk, J.; Leibschang, J.

1967-01-01

Most recent methods of tissue preservation are based on lyophilization and sterilization with gamma rays. Unfortunately the tissues preserved by this method lose their viability, this being connected with protein denaturation. The denaturing influence either of lyophilization or sterilization with gamma rays on different materials has been described. However, no observations on denaturation of proteins in prepared grafts are known. The work aimed at establishing the influence of individual stages of the procedure used in a tissue bank on the solubility of proteins. An experiment was performed using rat liver as a model tissue. Solubility of protein was determined in five groups of material, as follows: (1) fresh tissue, used as a control, (2) frozen tissue, (3) frozen and lyophilized tissue, (4) frozen, lyophilized and irradiated tissue, and (5) fresh irradiated tissue. Folin's method was used for determination of protein in water extracts of tissues. It was found that: (1) the whole procedure considerably diminished the protein solubility, (2) freezing diminishes the protein solubility by 35% on average, (3) lyophilization causes no further protein denaturation, (4) protein solubility is reduced most (by about 65%) by sterilization with gamma rays. (author)

High-level expression of soluble recombinant proteins in Escherichia coli using an HE-maltotriose-binding protein fusion tag.

Science.gov (United States)

Han, Yingqian; Guo, Wanying; Su, Bingqian; Guo, Yujie; Wang, Jiang; Chu, Beibei; Yang, Guoyu

2018-02-01

Recombinant proteins are commonly expressed in prokaryotic expression systems for large-scale production. The use of genetically engineered affinity and solubility enhancing fusion proteins has increased greatly in recent years, and there now exists a considerable repertoire of these that can be used to enhance the expression, stability, solubility, folding, and purification of their fusion partner. Here, a modified histidine tag (HE) used as an affinity tag was employed together with a truncated maltotriose-binding protein (MBP; consisting of residues 59-433) from Pyrococcus furiosus as a solubility enhancing tag accompanying a tobacco etch virus protease-recognition site for protein expression and purification in Escherichia coli. Various proteins tagged at the N-terminus with HE-MBP(Pyr) were expressed in E. coli BL21(DE3) cells to determine expression and solubility relative to those tagged with His6-MBP or His6-MBP(Pyr). Furthermore, four HE-MBP(Pyr)-fused proteins were purified by immobilized metal affinity chromatography to assess the affinity of HE with immobilized Ni 2+ . Our results showed that HE-MBP(Pyr) represents an attractive fusion protein allowing high levels of soluble expression and purification of recombinant protein in E. coli. Copyright © 2017 Elsevier Inc. All rights reserved.

Efficient protein production method for NMR using soluble protein tags with cold shock expression vector

Energy Technology Data Exchange (ETDEWEB)

Hayashi, Kokoro [Fujifilm Corporation, Analysis Technology Center (Japan); Kojima, Chojiro, E-mail: kojima@protein.osaka-u.ac.j [Nara Institute of Science and Technology (NAIST), Graduate School of Biological Sciences (Japan)

2010-11-15

The E. coli protein expression system is one of the most useful methods employed for NMR sample preparation. However, the production of some recombinant proteins in E. coli is often hampered by difficulties such as low expression level and low solubility. To address these problems, a modified cold-shock expression system containing a glutathione S-transferase (GST) tag, the pCold-GST system, was investigated. The pCold-GST system successfully expressed 9 out of 10 proteins that otherwise could not be expressed using a conventional E. coli expression system. Here, we applied the pCold-GST system to 84 proteins and 78 proteins were successfully expressed in the soluble fraction. Three other cold-shock expression systems containing a maltose binding protein tag (pCold-MBP), protein G B1 domain tag (pCold-GB1) or thioredoxin tag (pCold-Trx) were also developed to improve the yield. Additionally, we show that a C-terminal proline tag, which is invisible in {sup 1}H-{sup 15}N HSQC spectra, inhibits protein degradation and increases the final yield of unstable proteins. The purified proteins were amenable to NMR analyses. These data suggest that pCold expression systems combined with soluble protein tags can be utilized to improve the expression and purification of various proteins for NMR analysis.

BRI1 and BAK1 interact with G proteins and regulate sugar-responsive growth and development in Arabidopsis.

Science.gov (United States)

Peng, Yuancheng; Chen, Liangliang; Li, Shengjun; Zhang, Yueying; Xu, Ran; Liu, Zupei; Liu, Wuxia; Kong, Jingjing; Huang, Xiahe; Wang, Yingchun; Cheng, Beijiu; Zheng, Leiying; Li, Yunhai

2018-04-18

Sugars function as signal molecules to regulate growth, development, and gene expression in plants, yeasts, and animals. A coordination of sugar availability with phytohormone signals is crucial for plant growth and development. The molecular link between sugar availability and hormone-dependent plant growth are largely unknown. Here we report that BRI1 and BAK1 are involved in sugar-responsive growth and development. Glucose influences the physical interactions and phosphorylations of BRI1 and BAK1 in a concentration-dependent manner. BRI1 and BAK1 physically interact with G proteins that are essential for mediating sugar signaling. Biochemical data show that BRI1 can phosphorylate G protein β subunit and γ subunits, and BAK1 can phosphorylate G protein γ subunits. Genetic analyses suggest that BRI1 and BAK1 function in a common pathway with G-protein subunits to regulate sugar responses. Thus, our findings reveal an important genetic and molecular mechanism by which BR receptors associate with G proteins to regulate sugar-responsive growth and development.

Motile hepatocellular carcinoma cells preferentially secret sugar metabolism regulatory proteins via exosomes.

Science.gov (United States)

Zhang, Jing; Lu, Shaohua; Zhou, Ye; Meng, Kun; Chen, Zhipeng; Cui, Yizhi; Shi, Yunfeng; Wang, Tong; He, Qing-Yu

2017-07-01

Exosomes are deliverers of critically functional proteins, capable of transforming target cells in numerous cancers, including hepatocellular carcinoma (HCC). We hypothesize that the motility of HCC cells can be featured by comparative proteome of exosomes. Hence, we performed the super-SILAC-based MS analysis on the exosomes secreted by three human HCC cell lines, including the non-motile Hep3B cell, and the motile 97H and LM3 cells. More than 1400 exosomal proteins were confidently quantified in each MS analysis with highly biological reproducibility. We justified that 469 and 443 exosomal proteins represented differentially expressed proteins (DEPs) in the 97H/Hep3B and LM3/Hep3B comparisons, respectively. These DEPs focused on sugar metabolism-centric canonical pathways per ingenuity pathway analysis, which was consistent with the gene ontology analysis on biological process enrichment. These pathways included glycolysis I, gluconeogenesis I and pentose phosphate pathways; and the DEPs enriched in these pathways could form a tightly connected network. By analyzing the relative abundance of proteins and translating mRNAs, we found significantly positive correlation between exosomes and cells. The involved exosomal proteins were again focusing on sugar metabolism. In conclusion, motile HCC cells tend to preferentially export more sugar metabolism-associated proteins via exosomes that differentiate them from non-motile HCC cells. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Ageing mechanisms in chickpea seeds: Relationship of sugar hydrolysis and lipid peroxidation with Amadori and Millard reactions

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mahdi shaaban

2017-05-01

Full Text Available This experiment was performed in order to study on ageing mechanisms of chickpea seeds (Cicer arietinum L. in natural storage and accelerated ageing conditions in seed laboratory of Gorgan Agricultural Science and Natural Resources, Gorgan, Iran at 2015. Experiment was in completely randomized design arrangement with four replications. Treatments were 2 and 4 years natural storage and 1-5 days of accelerated ageing with control treatment. The results showed that with increasing of natural storage and accelerated ageing duration, germination percentage was decreased. Increasing of ageing duration decreased soluble sugars, non-reducing sugars and soluble proteins but lipid peroxidation, reducing sugars, protein carbonylation and Amadori and Millard reaction were increased. In natural storage condition lipid peroxidation was more than sugar hydrolysis but in accelerated ageing condition sugar hydrolysis was more than lipid peroxidation. These results show that the main reason of Amadori and Millard reaction in chickpea seeds in natural storage condition is lipid peroxidation and in accelerated ageing condition is sugar hydrolysis. Also, the results showed that Amadori reaction in natural storage condition was more than Amadori reaction and in accelerated ageing condition Millard reaction was more than Amadori reaction. The results of the present study showed that sever Millard reaction after Amadori reaction induced higher damage on seed and results to more decrease of seed viability and reduce of seed germination percentage in accelerated ageing than natural storage.

Evaluation of proline, chlorophyll, soluble sugar content and uptake of nutrients in the German chamomile (Matricaria chamomilla L. under drought stress and organic fertilizer treatments

Directory of Open Access Journals (Sweden)

Amin Salehi

2016-10-01

Conclusions: Totally, organic fertilization by vermicompost could partly alleviate the effect of drought stress on chamomile by increasing N, P and K uptake and leaf soluble sugar, especially in stressed treatments.

Cold-season patterns of reserve and soluble carbohydrates in sugar maple and ice-damaged trees of two age classes following drought

Science.gov (United States)

B. L. Wong; K. L. Baggett; A. H. Rye

2009-01-01

This study examines the effects of summer drought on the composition and profiles of cold-season reserve and soluble carbohydrates in sugar maple (Acer saccharum Marsh.) trees (50-100 years old or ~200 years old) in which the crowns were nondamaged or damaged by the 1998 ice storm. The overall cold season reserve...

Light exposure during storage preserving soluble sugar and l-ascorbic acid content of minimally processed romaine lettuce (Lactuca sativa L.var. longifolia).

Science.gov (United States)

Zhan, Lijuan; Hu, Jinqiang; Ai, Zhilu; Pang, Lingyun; Li, Yu; Zhu, Meiyun

2013-01-01

Minimally processed romaine lettuce (MPRL) leaves were stored in light condition (2500lux) or darkness at 4°C for 7d. Light exposure significantly delayed the degradation of chlorophyll and decrease of glucose, reducing sugar, and sucrose content, and thus preserved more total soluble solid (TSS) content at the end of storage in comparison with darkness. While, it did not influenced starch content that progressively decreased over time. The l-ascorbic acid (AA) accumulated in light-stored leaves, but deteriorated in dark-stored leaves during storage. The dehydroascorbic acid (DHA) increased in all leaves stored in both light and dark condition, of which light condition resulted in less DHA than darkness. In addition, the fresh weight loss and dry matter significantly increased and these increases were accelerated by light exposure. Conclusively, light exposure in applied intensity effectively alleviated MPRL quality deterioration by delaying the decreases of pigments, soluble sugar, TSS content and accumulating AA. Copyright © 2012 Elsevier Ltd. All rights reserved.

Lipid and sugar profiles of various barley cultivars (Hordeum vulgare

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Pastor Kristian A.

2015-01-01

Full Text Available The lipid components and soluble sugars in flour samples of different cultivars of barley (Hordeum vulgare, involving winter malting barley, winter forage barley, spring barley, and hulless barley, were identified. Fatty acids were extracted from flour samples with n-hexane, and derivatized into volatile methyl esters, using TMSH (trimethylsulfonium hydroxide in methanol. Soluble sugars were extracted from defatted and dried samples of barley flour with 96% ethanol, and further derivatized into the corresponding trimethylsilyl (TMS oximes, using hydroxylamine hydrochloride solution and BSTFA (N,O-bis-(trimethylsilyl-trifluoroacetamide. The hexane and alcoholic extracts of barley cultivars were analyzed by GC-MS system. Lipid and sugar compositions were very similar in all barley cultivars. Therefore, multivariate analysis was applied to numerical values of automatically integrated areas of the identified fatty acid methyl esters and TMS oximes of soluble sugars. The application of hierarchical cluster analysis showed a great similarity between the investigated flour samples of barley cultivars, according to their fatty acid content (0.96. Also, significant, but somewhat less similarity was observed regarding the content of soluble sugars (0.70. These preliminary results indicate the possibility of distinguishing flour made of barley, regardless of the variety, from flours made of other cereal species, just by the analysis of the contents of fatty acids and soluble sugars.[Projekat Ministarstva nauke Republike Srbije, br. TR 31066

Soluble Non-ammonia Nitrogen in Ruminal and Omasal Digesta of Korean Native Steers Supplemented with Soluble Proteins

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C. W. Choi

2012-09-01

Full Text Available An experiment was conducted to study the effect of soluble protein supplements on concentration of soluble non-ammonia nitrogen (SNAN in the liquid phase of ruminal (RD and omasal digesta (OD of Korean native steers, and to investigate diurnal pattern in SNAN concentration in RD and OD. Three ruminally cannulated Korean native steers in a 3×3 Latin square design consumed a basal diet of rice straw and corn-based concentrate (control, and that supplemented (kg/d DM basis with intact casein (0.24; IC or acid hydrolyzed casein (0.46; AHC. Ruminal digesta was sampled using a vacuum pump, whereas OD was collected using an omasal sampling system at 2.0 h intervals after a morning feeding. The SNAN fractions (free amino acid (AA, peptide and soluble protein in RD and OD were assessed using the ninhydrin assay. Concentrations of free AA and total SNAN in RD were significantly (p<0.05 lower than those in OD. Although free AA concentration was relatively high, mean peptide was quantitatively the most important fraction of total SNAN in both RD and OD, indicating that degradation of peptide to AA rather than hydrolysis of soluble protein to peptide or deamination may be the most limiting step in rumen proteolysis of Korean native steers. Diurnal variation in peptide concentration in OD for the soluble protein supplemented diets during the feeding cycle peaked 2 h post-feeding and decreased thereafter whereas that for the control was relatively constant during the entire feeding cycle. Diurnal variation in peptide concentration was rather similar between RD and OD.

Soluble Sugars as the Carbohydrate Reserve for CAM in Pineapple Leaves : Implications for the Role of Pyrophosphate:6-Phosphofructokinase in Glycolysis.

Science.gov (United States)

Carnal, N W; Black, C C

1989-05-01

Neutral ethanol-soluble sugar pools serve as carbohydrate reserves for Crassulacean acid metabolism (CAM) in pineapple (Ananas comosus (L.) Merr.) leaves. Levels of neutral soluble sugars and glucans fluctuated reciprocally with concentrations of malic acid. Hexose loss from neutral soluble-sugar pools was sufficient to account for malic acid accumulation with about 95% of the required hexose accounted for by turnover of fructose and glucose pools. Hexose loss from starch or starch plus lower molecular weight glucan pools was insufficient to account for nocturnal accumulation of malic acid. The apparent maximum catalytic capacity of pyrophosphate:6-phosphofructokinase (PPi-PFK) at 15 degrees C was about 16 times higher than the mean maximum rate of glycolysis that occurred to support malic acid accumulation in pineapple leaves at night and 12 times higher than the mean maximum rate of hexose turnover from all carbohydrate pools. The apparent maximum catalytic capacity of ATP-PFK at 15 degrees C was about 70% of the activity required to account for the mean maximal rate of hexose turnover from all carbohydrate pools if turnover were completely via glycolysis, and marginally sufficient to account for mean maximal rates of acidification. Therefore, at low night temperatures conducive to CAM and under subsaturating substrate concentrations, PPi-PFK activity, but not ATP-PFK activity, would be sufficient to support the rate of glycolytic carbohydrate processing required for acid accumulation. These data for pineapple establish that there are at least two types of CAM plants with respect to the nature of the carbohydrate reserve utilized to support nighttime CO(2) accumulation. The data further indicate that the glycolytic carbohydrate processing that supports acidification proceeds in different subcellular compartments in plants utilizing different carbohydrate reserves.

Novel mitochondria-targeted heat-soluble proteins identified in the anhydrobiotic Tardigrade improve osmotic tolerance of human cells.

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Sae Tanaka

Full Text Available Tardigrades are able to tolerate almost complete dehydration through transition to a metabolically inactive state, called "anhydrobiosis". Late Embryogenesis Abundant (LEA proteins are heat-soluble proteins involved in the desiccation tolerance of many anhydrobiotic organisms. Tardigrades, Ramazzottius varieornatus, however, express predominantly tardigrade-unique heat-soluble proteins: CAHS (Cytoplasmic Abundant Heat Soluble and SAHS (Secretory Abundant Heat Soluble proteins, which are secreted or localized in most intracellular compartments, except the mitochondria. Although mitochondrial integrity is crucial to ensure cellular survival, protective molecules for mitochondria have remained elusive. Here, we identified two novel mitochondrial heat-soluble proteins, RvLEAM and MAHS (Mitochondrial Abundant Heat Soluble, as potent mitochondrial protectants from Ramazzottius varieornatus. RvLEAM is a group3 LEA protein and immunohistochemistry confirmed its mitochondrial localization in tardigrade cells. MAHS-green fluorescent protein fusion protein localized in human mitochondria and was heat-soluble in vitro, though no sequence similarity with other known proteins was found, and one region was conserved among tardigrades. Furthermore, we demonstrated that RvLEAM protein as well as MAHS protein improved the hyperosmotic tolerance of human cells. The findings of the present study revealed that tardigrade mitochondria contain at least two types of heat-soluble proteins that might have protective roles in water-deficient environments.

Leaching behavior of water-soluble carbohydrates from almond hulls

Science.gov (United States)

Over 58% of the dry matter content of the hulls from the commercial almond (Prunus dulcis (Miller) D.A. Webb) is soluble in warm water (50-70°C) extraction. The water-soluble extractables include useful amounts of fermentable sugars (glucose, fructose, sucrose), sugar alcohols (inositol and sorbito...

Urinary Excretion of Sodium, Nitrogen, and Sugar Amounts Are Valid Biomarkers of Dietary Sodium, Protein, and High Sugar Intake in Nonobese Adolescents.

Science.gov (United States)

Moore, Lori B; Liu, Sarah V; Halliday, Tanya M; Neilson, Andrew P; Hedrick, Valisa E; Davy, Brenda M

2017-12-01

Background: Objective indicators of dietary intake (e.g., biomarkers) are needed to overcome the limitations of self-reported dietary intake assessment methods in adolescents. To our knowledge, no controlled feeding studies to date have evaluated the validity of urinary sodium, nitrogen, or sugar excretion as dietary biomarkers in adolescents. Objective: This investigation aimed to evaluate the validity of urinary sodium, nitrogen, and total sugars (TS) excretion as biomarkers for sodium, protein, and added sugars (AS) intake in nonobese adolescents. Methods: In a crossover controlled feeding study design, 33 adolescents [12-18 y of age, 47 ± 25th percentile (mean ± SD) of body mass index (BMI; in kg/m 2 ) for age] consumed 5% AS [low added sugars (LAS)] and 25% AS [high added sugars (HAS)] isocaloric, macronutrient-matched (55% carbohydrate, 30% fat, and 15% protein) diets for 7 d each, in a randomly assigned order, with a 4-wk washout period between diets. On the final 2 d of each diet period, 24-h urine samples were collected. Thirty-two adolescents completed all measurements (97% retention). Results: Urinary sodium was not different from the expected 90% recovery (mean ± SD: 88% ± 18%, P = 0.50). Urinary nitrogen was correlated with protein intake ( r = 0.69, P sodium appears to be a valid biomarker for sodium intake in nonobese adolescents. Urinary nitrogen is associated with protein intake, but nitrogen excretion rates were less than previously reported for adults, possibly owing to adolescent growth rates. TS excretion reflects AS at 25% AS intake and was responsive to the change in AS intake. Thus, urinary biomarkers are promising objective indicators of dietary intake in adolescents, although larger-scale feeding trials are needed to confirm these findings. This trial was registered at clinicaltrials.gov as NCT02455388. © 2017 American Society for Nutrition.

Water-Deficit Tolerance in Sweet Potato [Ipomoea batatas (L. Lam.] by Foliar Application of Paclobutrazol: Role of Soluble Sugar and Free Proline

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Suravoot Yooyongwech

2017-08-01

Full Text Available The objective of this study was to elevate water deficit tolerance by improving soluble sugar and free proline accumulation, photosynthetic pigment stabilization, photosynthetic abilities, growth performance and storage root yield in sweet potato cv. ‘Tainung 57’ using a foliar application of paclobutrazol (PBZ. The experiment followed a Completely Randomized Block Design with four concentrations of PBZ: 0 (control, 17, 34, and 51 μM before exposure to 47.5% (well irrigation, 32.3% (mild water deficit or 17.5% (severe water deficit soil water content. A sweet potato cultivar, ‘Japanese Yellow’, with water deficit tolerance attributes was the positive check in this study. Total soluble sugar content (sucrose, glucose, and fructose increased by 3.96-folds in ‘Tainung 57’ plants treated with 34 μM PBZ grown under 32.3% soil water content (SWC compared to the untreated plants, adjusting osmotic potential in the leaves and controlling stomatal closure (represented by stomatal conductance and transpiration rate. In addition, under the same treatment, free proline content (2.15 μmol g-1 FW increased by 3.84-folds when exposed to 17.5% SWC. PBZ had an improved effect on leaf size, vine length, photosynthetic pigment stability, chlorophyll fluorescence, and net photosynthetic rate; hence, delaying wilting symptoms and maintaining storage root yield (26.93 g plant-1 at the harvesting stage. A positive relationship between photon yield of PSII (ΦPSII and net photosynthetic rate was demonstrated (r2 = 0.73. The study concludes that soluble sugar and free proline enrichment in PBZ-pretreated plants may play a critical role as major osmoprotectant to control leaf osmotic potential and stomatal closure when plants were subjected to low soil water content, therefore, maintaining the physiological and morphological characters as well as storage root yield.

Water-Deficit Tolerance in Sweet Potato [Ipomoea batatas (L.) Lam.] by Foliar Application of Paclobutrazol: Role of Soluble Sugar and Free Proline.

Science.gov (United States)

Yooyongwech, Suravoot; Samphumphuang, Thapanee; Tisarum, Rujira; Theerawitaya, Cattarin; Cha-Um, Suriyan

2017-01-01

The objective of this study was to elevate water deficit tolerance by improving soluble sugar and free proline accumulation, photosynthetic pigment stabilization, photosynthetic abilities, growth performance and storage root yield in sweet potato cv. 'Tainung 57' using a foliar application of paclobutrazol (PBZ). The experiment followed a Completely Randomized Block Design with four concentrations of PBZ: 0 (control), 17, 34, and 51 μM before exposure to 47.5% (well irrigation), 32.3% (mild water deficit) or 17.5% (severe water deficit) soil water content. A sweet potato cultivar, 'Japanese Yellow', with water deficit tolerance attributes was the positive check in this study. Total soluble sugar content (sucrose, glucose, and fructose) increased by 3.96-folds in 'Tainung 57' plants treated with 34 μM PBZ grown under 32.3% soil water content (SWC) compared to the untreated plants, adjusting osmotic potential in the leaves and controlling stomatal closure (represented by stomatal conductance and transpiration rate). In addition, under the same treatment, free proline content (2.15 μmol g -1 FW) increased by 3.84-folds when exposed to 17.5% SWC. PBZ had an improved effect on leaf size, vine length, photosynthetic pigment stability, chlorophyll fluorescence, and net photosynthetic rate; hence, delaying wilting symptoms and maintaining storage root yield (26.93 g plant -1 ) at the harvesting stage. A positive relationship between photon yield of PSII (Φ PSII ) and net photosynthetic rate was demonstrated ( r 2 = 0.73). The study concludes that soluble sugar and free proline enrichment in PBZ-pretreated plants may play a critical role as major osmoprotectant to control leaf osmotic potential and stomatal closure when plants were subjected to low soil water content, therefore, maintaining the physiological and morphological characters as well as storage root yield.

Soluble protein isolated from low cost fish and fish wastes

OpenAIRE

Lekshmy Nair, A.; Gopakumar, K.

1982-01-01

The method of preparation, composition, amino acid content, protein efficiency ratio and areas of possible application of water soluble protein isolates from low cost fish and fish wastes are discussed in detail in this communication.

Effects of gamma irradiation on physicochemical properties of heat-induced gel prepared with chicken salt-soluble proteins

International Nuclear Information System (INIS)

Choi, Yun-Sang; Kim, Hyun-Wook; Hwang, Ko-Eun; Song, Dong-Heon; Jeong, Tae-Jun; Seo, Kwang-Wook; Kim, Young-Boong; Kim, Cheon-Jei

2015-01-01

The technological effects of gamma irradiation (0, 3, 7, and 10 kGy) on chicken salt-soluble meat proteins in a model system were investigated. There were no significant differences in protein, fat, and ash content, and sarcoplasmic protein solubility among all samples. The samples with increasing gamma irradiation levels had higher pH, lightness, yellowness, and apparent viscosity, whereas moisture content, water holding capacity, redness, myofibrillar protein solubility, total protein solubility, hardness, springiness, cohesiveness, gumminess, and chewiness were the highest in the unirradiated control. The result from meat products using gamma irradiation was intended to provide a basic resource processing technology. - Highlights: • The effect of gamma irradiation on salt-soluble meat proteins was investigated. • Gelling properties of salt-soluble protein affected by gamma irradiation. • Gamma irradiation of meat products provides a basic resource processing technology

Arabidopsis scaffold protein RACK1A modulates rare sugar D-allose regulated gibberellin signaling

OpenAIRE

Fennell, Herman; Olawin, Abdulquadri; Mizanur, Rahman M.; Izumori, Ken; Chen, Jin-Gui; Ullah, Hemayet

2012-01-01

As energy sources and structural components, sugars are the central regulators of plant growth and development. In addition to the abundant natural sugars in plants, more than 50 different kinds of rare sugars exist in nature, several of which show distinct roles in plant growth and development. Recently, one of the rare sugars, D-allose, an epimer of D-glucose at C3, is found to suppress plant hormone gibberellin (GA) signaling in rice. Scaffold protein RACK1A in the model plant Arabidopsis ...

Changes in protein solubility, fermentative capacity, viscoelasticity ...

African Journals Online (AJOL)

Frozen dough should be stored for fewer than 21 days; time in which the loaf volume of bread made from frozen dough was approximately 40.84% smaller than that of fresh bread dough formulation. Keywords: French type bread, frozen dough, protein solubility, baking quality, viscoelasticity. African Journal of Biotechnology ...

The Changes in Color, Soluble Sugars, Organic Acids, Anthocyanins and Aroma Components in “Starkrimson” during the Ripening Period in China

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Yulian Liu

2016-06-01

Full Text Available “Starkrimson” is a traditional apple cultivar that was developed a long time ago and was widely cultivated in the arid region of the northern Wei River of China. However, little information regarding the quality characteristics of “Starkrimson” fruit has been reported in this area. To elucidate these characteristics, the color, soluble sugars, organic acids, anthocyanins and aroma components were measured during the ripening period through the use of high performance liquid chromatography (HPLC and gas chromatography-mass spectrometry (GC-MS. The results indicated that the changes in anthocyanin contents took place later than the changes in the Commission International Eclairage (CIE parameters. Meanwhile, cyanidin 3-galactoside (cy3-gal, fructose, sucrose, glucose and malic acid were the primary organic compounds, and 1-butanol-2-methyl-acetate, 2-hexenal and 1-hexanol were the most abundant aroma components in the skin. Furthermore, rapidly changing soluble sugars and organic acid synchronization took place in the early ripening period, while rapidly changing aroma components occurred later, on the basis of fresh weight. This result suggested that the production of aroma components might be a useful index of apple maturity.

Influence of cold stress on contents of soluble sugars, vitamin C and free amino acids including gamma-aminobutyric acid (GABA) in spinach (Spinacia oleracea).

Science.gov (United States)

Yoon, Young-Eun; Kuppusamy, Saranya; Cho, Kye Man; Kim, Pil Joo; Kwack, Yong-Bum; Lee, Yong Bok

2017-01-15

The contents of soluble sugars (sucrose, fructose, glucose, maltose and raffinose), vitamin C and free amino acids (34 compounds, essential and non-essential) were quantified in open-field and greenhouse-grown spinaches in response to cold stress using liquid chromatography. In general, greenhouse cultivation produced nutritionally high value spinach in a shorter growing period, where the soluble sugars, vitamin C and total amino acids concentrations, including essential were in larger amounts compared to those grown in open-field scenarios. Further, low temperature exposure of spinach during a shorter growth period resulted in the production of spinach with high sucrose, ascorbate, proline, gamma-aminobutyric acid, valine and leucine content, and these constitute the most important energy/nutrient sources. In conclusion, cultivation of spinach in greenhouse at a low temperature (4-7°C) and exposure for a shorter period (7-21days) before harvest is recommended. This strategy will produce a high quality product that people can eat. Copyright © 2016 Elsevier Ltd. All rights reserved.

Growth responses and accumulation of soluble sugars in Inga marginata Wild. (Fabaceae) subjected to flooding under contrasting light conditions.

Science.gov (United States)

Bender, B; Capellesso, E S; Lottici, M E; Sentkovski, J; Mielniczki-Pereira, A A; Rosa, L M G; Sausen, T L

2017-01-01

Flood events in riparian forests of southern Brazil, can be characterized as unpredictable and of low magnitude with an average duration of less than 15 days. Inga marginata is an evergreen tree which grows in Southeast South America on a wide range of environments, including riparian forests. In this paper, the interactive effects of the light environment and soil flooding on morphological parameters of I. marginata were examined. Seedlings were acclimated in two contrasting light conditions: sun or shade for 30 days. Sun and shade plants were subjected to soil flooding for two periods; five or 15 days. After 5 days, the interaction between flooding and light did not affect growth, chlorophyll content and dry mass or the root-shoot ratio. After 15 days, flooded plants from the sunny treatment had a lower shoot dry mass compared to control sun plants and flooded plants from the shaded treatment. Moreover, the higher dry mass observed for shade plants compared to sun plants, following flooding, can also be directly associated with a higher content of soluble sugars. Shade plants of I. marginata showed a greater acclimation to soil waterlogging. This acclimation appears to be associated with a larger accumulation of soluble sugars compared to non-flooded plants. The responses observed on the shade plants appear to be decisive to indicate the use of I. marginata in degraded areas.

Effect of gamma irradiation on the sugar and protein composition of Iraqi dates

International Nuclear Information System (INIS)

Auda, H.; Khalaf, Z.; Mirjan, J.

1978-01-01

Completely ripened date fruits of several Iraqi varieties were irradiated with 30, 70, 100, 270 and 500krad of gamma irradiation and stored at a temperature of 25-35 0 C in wooden boxes or in plastic bags. At various intervals samples were taken and analysed by paper-chromatography, gas-chromatography and spectrophotometry for quantitative and qualitative changes in carbohydrates. The results showed no effect of gamma irradiation on reducing sugar and major carbohydrate components. The formation of malonaldehyde under gamma irradiation of dates and solid standard sugars was also studied up to 500krad. The results showed no formation of malonaldehyde in irradiated date samples as well as standard sugars. Gamma irradiation showed no effect on the protein content of dates. However, storage showed some reduction in the protein content of both unirradiated and irradiated samples. (author)

Enantiomer Ratios of Meteoritic Sugar Derivatives

Science.gov (United States)

Cooper, George

2012-01-01

Carbonaceous meteorites contain a diverse suite of soluble organic compounds. Studies of these compounds reveal the Solar System's earliest organic chemistry. Among the classes of organic compounds found in meteorites are keto acids (pyruvic acid, etc.), hydroxy tricarboxylic acids (1), amino acids, amides, purines and pyrimidines. The Murchison and Murray meteorites are the most studied for soluble and insoluble organic compounds and organic carbon phases. The majority of (indigenous) meteoritic compounds are racemic, (i.e., their D/L enantiomer ratios are 50:50). However, some of the more unusual (non-protein) amino acids contain slightly more of one enantiomer (usually the L) than the other. This presentation focuses on the enantiomer analyses of three to six-carbon (3C to 6C) meteoritic sugar acids. The molecular and enantiomer analysis of corresponding sugar alcohols will also be discussed. Detailed analytical procedures for sugar-acid enantiomers have been described. Results of several meteorite analyses show that glyceric acid is consistently racemic (or nearly so) as expected of non-biological mechanisms of synthesis. Also racemic are 4-C deoxy sugar acids: 2-methyl glyceric acid; 2,4-dihydroxybutyric acid; 2,3-dihydroxybutyric acid (two diastereomers); and 3,4-dihydroxybutyric acid. However, a 4C acid, threonic acid, has never been observed as racemic, i.e., it possesses a large D excess. In several samples of Murchison and one of GRA 95229 (possibly the most pristine carbonaceous meteorite yet analyzed) threonic acid has nearly the same D enrichment. In Murchison, preliminary isotopic measurements of individual threonic acid enantiomers point towards extraterrestrial sources of the D enrichment. Enantiomer analyses of the 5C mono-sugar acids, ribonic, arabinonic, xylonic, and lyxonic also show large D excesses. It is worth noting that all four of these acids (all of the possible straight-chained 5C sugar acids) are present in meteorites, including the

Cell culture media supplementation of infrequently used sugars for the targeted shifting of protein glycosylation profiles.

Science.gov (United States)

Hossler, Patrick; Racicot, Christopher; Chumsae, Christopher; McDermott, Sean; Cochran, Keith

2017-03-01

Mammalian cells in culture rely on sources of carbohydrates to supply the energy requirements for proliferation. In addition, carbohydrates provide a large source of the carbon supply for supporting various other metabolic activities, including the intermediates involved in the protein glycosylation pathway. Glucose and galactose, in particular, are commonly used sugars in culture media for these purposes. However, there exists a very large repertoire of other sugars in nature, and many that have been chemically synthesized. These sugars are particularly interesting because they can be utilized by cells in culture in distinct ways. In the present work it has been found that many infrequently used sugars, and the corresponding cellular response towards them as substrates, led to differences in the protein N-glycosylation profile of a recombinant glycoprotein. The selective media supplementation of raffinose, trehalose, turanose, palatinose, melezitose, psicose, lactose, lactulose, and mannose were found to be capable of redirecting N-glycan oligosaccharide profiles. Despite this shifting of protein glycosylation, there were no other adverse changes in culture performance, including both cell growth and cellular productivity over a wide range of supplemented sugar concentrations. The approach presented highlights a potential means towards both the targeted shifting of protein glycosylation profiles and ensuring recombinant protein comparability, which up to this point in time has remained under-appreciated for these under-utilized compounds. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:511-522, 2017. © 2017 American Institute of Chemical Engineers.

Effects of sugar alcohol and proteins on the survival of Lactobacillus bulgaricus LB6 during freeze drying.

Science.gov (United States)

Chen, He; Chen, Shiwei; Chen, Hongli; Wu, Yanyan; Shu, Guowei

2015-01-01

Lactobacillus bulgaricus LB6 is a bacterium which was selected in the commercial yoghurt with high angiotensin converting enzyme (ACE) inhibitory activity. Preparation of concentrated starter cultures via freeze drying is of practical importance to dairy and food industries. We optimized the optimal sugar alcohol and proteins for Lactobacillus bulgaricus LB6 during the process of freeze drying using a Plackett-Burman design. In our initial tests survival rate and the number of viable cells were associated with the type of lyoprotectant used and so our optimization protocol focused on increasing survival rate. Substances that had previously had a protective effect during freeze drying were investigated, for example: mannitol, sorbitol, xylitol, meso-erythritol, lactitol, whey protein isolate 90, bovine serum albumin, and whey protein concentrate 80 and soy protein isolate 70. We found that the optimum sugar alcohol and proteins for survival of Lactobacillus bulgaricus LB6 were whey protein concentrate (p = 0.0040 for survival rate), xylitol (p = 0.0067 for survival rate) and sorbitol (p = 0.0073 for survival rate), they showed positive effect (whey protein concentrate and sorbitol) or negative effect (xylitol). The effectiveness of three chosen sugar alcohols and protein implied that they could be used as lyoprotectant for Lactobacillus bulgaricus LB6 in the further research, the optimal composition of sugar alcohol and protein for the lyoprotectant use must be established.

Characterization of a new antifungal non-specific lipid transfer protein (nsLTP) from sugar beet leaves

DEFF Research Database (Denmark)

Kristensen, A K; Brunstedt, J; Madsen, M T

2000-01-01

A novel protein (IWF5) comprising 92 amino acids has been purified from the intercellular washing fluid of sugar beet leaves using cation exchange chromatography and reversed phase high performance liquid chromatography. Based on amino acid sequence homology, including the presence of eight...... cysteines at conserved positions, the protein can be classified as a member of the plant family of non-specific lipid transfer proteins (nsLTPs). The protein is 47% identical to IWF1, an antifungal nsLTP previously isolated from leaves of sugar beet. A potential site for N-linked glycosylation present...

A tonoplast sugar transporter underlies a sugar accumulation QTL in watermelon

Science.gov (United States)

The molecular mechanism controlling accumulation of soluble sugars in watermelon (Citrullus lanatus) fruit, a trait associated with sweet-dessert watermelon domestication, is still unknown. We re-sequenced 96 recombinant inbred lines, derived from a cross between sweet and unsweet watermelon accessi...

Soluble Milk Protein Supplementation with Moderate Physical Activity Improves Locomotion Function in Aging Rats.

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Aude Lafoux

Full Text Available Aging is associated with a loss of muscle mass and functional capacity. Present study was designed to compare the impact of specific dairy proteins on muscular function with or without a low-intensity physical activity program on a treadmill in an aged rat model. We investigated the effects of nutritional supplementation, five days a week over a 2-month period with a slow digestible protein, casein or fast digestible proteins, whey or soluble milk protein, on strength and locomotor parameters in sedentary or active aged Wistar RjHan rats (17-19 months of age. An extensive gait analysis was performed before and after protein supplementation. After two months of protein administration and activity program, muscle force was evaluated using a grip test, spontaneous activity using an open-field and muscular mass by specific muscle sampling. When aged rats were supplemented with proteins without exercise, only minor effects of different diets on muscle mass and locomotion were observed: higher muscle mass in the casein group and improvement of stride frequencies with soluble milk protein. By contrast, supplementation with soluble milk protein just after physical activity was more effective at improving overall skeletal muscle function in old rats compared to casein. For active old rats supplemented with soluble milk protein, an increase in locomotor activity in the open field and an enhancement of static and dynamic gait parameters compared to active groups supplemented with casein or whey were observed without any differences in muscle mass and forelimb strength. These results suggest that consumption of soluble milk protein as a bolus immediately after a low intensity physical activity may be a suitable nutritional intervention to prevent decline in locomotion in aged rats and strengthen the interest to analyze the longitudinal aspect of locomotion in aged rodents.

Soluble Milk Protein Supplementation with Moderate Physical Activity Improves Locomotion Function in Aging Rats.

Science.gov (United States)

Lafoux, Aude; Baudry, Charlotte; Bonhomme, Cécile; Le Ruyet, Pascale; Huchet, Corinne

2016-01-01

Aging is associated with a loss of muscle mass and functional capacity. Present study was designed to compare the impact of specific dairy proteins on muscular function with or without a low-intensity physical activity program on a treadmill in an aged rat model. We investigated the effects of nutritional supplementation, five days a week over a 2-month period with a slow digestible protein, casein or fast digestible proteins, whey or soluble milk protein, on strength and locomotor parameters in sedentary or active aged Wistar RjHan rats (17-19 months of age). An extensive gait analysis was performed before and after protein supplementation. After two months of protein administration and activity program, muscle force was evaluated using a grip test, spontaneous activity using an open-field and muscular mass by specific muscle sampling. When aged rats were supplemented with proteins without exercise, only minor effects of different diets on muscle mass and locomotion were observed: higher muscle mass in the casein group and improvement of stride frequencies with soluble milk protein. By contrast, supplementation with soluble milk protein just after physical activity was more effective at improving overall skeletal muscle function in old rats compared to casein. For active old rats supplemented with soluble milk protein, an increase in locomotor activity in the open field and an enhancement of static and dynamic gait parameters compared to active groups supplemented with casein or whey were observed without any differences in muscle mass and forelimb strength. These results suggest that consumption of soluble milk protein as a bolus immediately after a low intensity physical activity may be a suitable nutritional intervention to prevent decline in locomotion in aged rats and strengthen the interest to analyze the longitudinal aspect of locomotion in aged rodents.

Production of bacterial protein from sugar cane bagasse pith

Energy Technology Data Exchange (ETDEWEB)

Molina, O E; Callieri, D A.S.; Perotti de Galvez, N

1980-01-01

Bacterial protein was produced during the fermentation of sugar cane bagasse pith (BP) by a mixture of cellulolytic bacteria, one of them being a species of Cellulomonas. If the BP were treated with 1% NaOH prior to fermentation, the liquor could be used twice more without affecting the yield of bacterial protein. After that, the liquor became too dark and impaired the subsequent washing of BP. If the concentration of N (as NaN0/sub 3/) in the fermentation medium were raised, the conversion factor to protein was lowered, but the amount of protein formed per L per h and the ratio of protein to BP became higher. The evolution of pH, the dry matter content, cellulolytic activity, and protein yield were all affected by the type of N source used. The yield of bacterial protein can probably be increased by automatically controlling the pH and dissolved O levels of the culture.

Sugars and Desiccation Tolerance in Seeds 1

Science.gov (United States)

Koster, Karen L.; Leopold, A. Carl

1988-01-01

Soluble sugars have been shown to protect liposomes and lobster microsomes from desiccation damage, and a protective role has been proposed for them in several anhydrous systems. We have studied the relationship between soluble sugar content and the loss of desiccation tolerance in the axes of germinating soybean (Glycine max L. Merr. cv Williams), pea (Pisum sativum L. cv Alaska), and corn (Zea mays L. cv Merit) axes. The loss of desiccation tolerance during imbibition was monitored by following the ability of seeds to germinate after desiccation following various periods of preimbibition and by following the rates of electrolyte leakage from dried, then rehydrated axes. Finally, we analyzed the soluble sugar contents of the axes throughout the transition from desiccation tolerance to intolerance. These analyses show that sucrose and larger oligosaccharides were consistently present during the tolerant stage, and that desiccation tolerance disappeared as the oligosaccharides were lost. The results support the idea that sucrose may serve as the principal agent of desiccation tolerance in these seeds, with the larger oligosaccharides serving to keep the sucrose from crystallizing. PMID:16666392

Sugar-starvation-induced changes of carbon metabolism in excised maize root tips

International Nuclear Information System (INIS)

Dieuaide-Noubhani, M.; Canioni, P.; Raymond, P.

1997-01-01

Excised maize (Zea mays L.) root tips were used to study the early metabolic effects of glucose (Glc) starvation. Root tips were prelabeled with [1-13C]Glc so that carbohydrates and metabolic intermediates were close to steady-state labeling, but lipids and proteins were scarcely labeled. They were then incubated in a sugar-deprived medium for carbon starvation. Changes in the level of soluble sugars, the respiratory quotient, and the 13C enrichment of intermediates, as measured by 13C and 1H nuclear magnetic resonance, were studied to detect changes in carbon fluxes through glycolysis and the tricarboxylic acid cycle. Labeling of glutamate carbons revealed two major changes in carbon input into the tricarboxylic acid cycle: (a) the phosphoenolpyruvate carboxylase flux stopped early after the start of Glc starvation, and (b) the contribution of glycolysis as the source of acetyl-coenzyme A for respiration decreased progressively, indicating an increasing contribution of the catabolism of protein amino acids, fatty acids, or both. The enrichment of glutamate carbons gave no evidence for proteolysis in the early steps of starvation, indicating that the catabolism of proteins was delayed compared with that of fatty acids. Labeling of carbohydrates showed that sucrose turnover continues during sugar starvation, but gave no indication for any significant flux through gluconeogenesis

Increase The Sugar Concentration of The Solution Sugar by Reverse Osmotic Membrane

Science.gov (United States)

Redjeki, S.; Hapsari, N.; Iriani

2018-01-01

Sugar is one of the basic needs of people and food and drink industry. As technology advances and the demand for efficient usage of sugar rises, crystal sugar is seen as less advantageous than liquid sugar. If sugar is always dissolved in water before use, then it will be more efficient and practical for consumers to use sugar in liquid form than in crystal form. Other than that, liquid sugar is also attractive to consumers because it is economical, hygienic, instantly soluble in hot and cold water, fresher and longer-lasting, able to thicken and enrich the texture of foods and drinks, and functions as sweetener, syrup, and flavor enhancer. Liquid sugar is also more beneficial for sugar producers because of simpler production process, cheaper production cost, and similar yield with no extra cost. In sugar production, separation process is found in most of its stages and therefore the use of membrane technology for separating solute and water content has a good potential. In this research, water content reduction of sugar solution was done in order to increase the sugar concentration of the solution. The parameters of this research were 4%, 5%, and 6% starting concentration of sugar solution; 20, 40, and 60 minutes of process time; and 85 and 60 PSI ΔP. The best result was acquired on 4% starting concentration, 60 PSI ΔP, and 60 minutes process time.

Screening of genetic parameters for soluble protein expression in Escherichia coli

DEFF Research Database (Denmark)

Vernet, Erik; Kotzsch, Alexander; Voldborg, Bjørn

2011-01-01

Soluble expression of proteins in a relevant form for functional and structural investigations still often remains a challenge. Although many biochemical factors are known to affect solubility, a thorough investigation of yield-limiting factors is normally not feasible in high-throughput efforts....... Here we present a screening strategy for expression of biomedically relevant proteins in Escherichia coli using a panel of six different genetic variations. These include engineered strains for rare codon supplementation, increased disulfide bond formation in the cytoplasm and novel vectors...... for secretion to the periplasm or culture medium. Combining these variants with expression construct truncations design, we report on parallel cloning and expression of more than 300 constructs representing 24 selected proteins; including full-length variants of human growth factors, interleukins and growth...

Immunochemical analyses of soluble lens proteins in some marine fishes

Digital Repository Service at National Institute of Oceanography (India)

Menezes, M.R.

Soluble eye lens proteins of 10 fishes, belonging to the families Clupeidae, Hemirhamphidae, Lactaridae, Scombridae, Stromatidae, Psettodidae, Bothidae and Soleidae were studied by immunoelectrophoresis using the lens antiserum of Sardinella...

Production and characterization of cowpea protein hydrolysate with optimum nitrogen solubility by enzymatic hydrolysis using pepsin.

Science.gov (United States)

Mune Mune, Martin Alain; Minka, Samuel René

2017-06-01

Cowpea is a source of low-cost and good nutritional quality protein for utilization in food formulations in replacement of animal proteins. Therefore it is necessary that cowpea protein exhibits good functionality, particularly protein solubility which affects the other functional properties. The objective of this study was to produce cowpea protein hydrolysate exhibiting optimum solubility by the adequate combination of hydrolysis parameters, namely time, solid/liquid ratio (SLR) and enzyme/substrate ratio (ESR), and to determine its functional properties and molecular characteristics. A Box-Behnken experimental design was used for the experiments, and a second-order polynomial to model the effects of hydrolysis time, SLR and ESR on the degree of hydrolysis and nitrogen solubility index. The optimum hydrolysis conditions of time 208.61 min, SLR 1/15 (w/w) and ESR 2.25% (w/w) yielded a nitrogen solubility of 75.71%. Protein breakdown and the peptide profile following enzymatic hydrolysis were evaluated by sodium dodecyl sulfate polyacrylamide gel electrophoresis and size exclusion chromatography. Cowpea protein hydrolysate showed higher oil absorption capacity, emulsifying activity and foaming ability compared with the concentrate. The solubility of cowpea protein hydrolysate was adequately optimized by response surface methodology, and the hydrolysate showed adequate functionality for use in food. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

High-Pressure-High-Temperature Processing Reduces Maillard Reaction and Viscosity in Whey Protein-Sugar Solutions.

Science.gov (United States)

Avila Ruiz, Geraldine; Xi, Bingyan; Minor, Marcel; Sala, Guido; van Boekel, Martinus; Fogliano, Vincenzo; Stieger, Markus

2016-09-28

The aim of the study was to determine the influence of pressure in high-pressure-high-temperature (HPHT) processing on Maillard reactions and protein aggregation of whey protein-sugar solutions. Solutions of whey protein isolate containing either glucose or trehalose at pH 6, 7, and 9 were treated by HPHT processing or conventional high-temperature (HT) treatments. Browning was reduced, and early and advanced Maillard reactions were retarded under HPHT processing at all pH values compared to HT treatment. HPHT induced a larger pH drop than HT treatments, especially at pH 9, which was not associated with Maillard reactions. After HPHT processing at pH 7, protein aggregation and viscosity of whey protein isolate-glucose/trehalose solutions remained unchanged. It was concluded that HPHT processing can potentially improve the quality of protein-sugar-containing foods, for which browning and high viscosities are undesired, such as high-protein beverages.

Biochemical characterization of soluble proteins in pecan [Carya illinoinensis (Wangenh.) K. Koch].

Science.gov (United States)

Venkatachalam, Mahesh; Roux, Kenneth H; Sathe, Shridhar K

2008-09-10

Pecans (cv. Desirable) contained approximately 10% protein on a dry weight basis. The minimum nitrogen solubility (5.9-7.5%) at 0.25-0.75 M trichloroacetic acid represented the nonprotein nitrogen. Among the solvents assessed for protein solubilization, 0.1 M NaOH was the most effective, while borate saline buffer (pH 8.45) was judged to be optimal for protein solubilization. The protein solubility was minimal in the pH range of 3-7 and significantly increased on either side of this pH range. Increasing the NaCl concentration from 0 to 4 M significantly improved ( approximately 8-fold increase) protein solubilization. Following Osborne protein fractionation, the alkali-soluble glutelin fraction (60.1%) accounted for a major portion of pecan proteins followed by globulin (31.5%), prolamin (3.4%), and albumin (1.5%), respectively. The majority of pecan polypeptides were in the molecular mass range of 12-66 kDa and in the pI range of 4.0-8.3. The pecan globulin fraction was characterized by the presence of several glycoprotein polypeptides. Lysine was the first limiting essential amino acid in the defatted flour, globulin, prolamin, and alkaline glutelin fractions. Leucine and tryptophan were the first limiting essential amino acids in albumin and acid glutelin fractions, respectively. Rabbit polyclonal antibodies detected a range of pecan polypeptides in the 12-60 kDa range, of which the globulin fraction contained the most reactive polypeptides.

Mobilization and Role of Starch, Protein, and Fat Reserves during Seed Germination of Six Wild Grassland Species.

Science.gov (United States)

Zhao, Ming; Zhang, Hongxiang; Yan, Hong; Qiu, Lu; Baskin, Carol C

2018-01-01

Since seed reserves can influence seed germination, the quantitative and qualitative differences in seed reserves may relate to the germination characteristics of species. The purpose of our study was to evaluate the correlation between germination and seed reserves, as well as their mobilization during germination of six grassland species ( Chloris virgata , Kochia scoparia , Lespedeza hedysaroides , Astragalus adsurgens , Leonurus artemisia , and Dracocephalum moldavica ) and compare the results with domesticated species. We measured starch, protein, and fat content in dry seeds and the initial absorption of water during imbibition. Starch, soluble protein, fat, and soluble sugar content also were determined at five stages during germination. Starch, protein, and fat reserves in dry seeds were not significantly correlated with germination percentage and rate (speed), but soluble sugar and soluble protein contents at different germination stages were positively significantly correlated with germination rate for the six species. Starch was mainly used during seed imbibition, and soluble protein was used from the imbibition stage to the highest germination stage. Fat content for all species remained relatively constant throughout germination for six species, regardless of the proportion of other seed reserves in the seeds. Our results for fat utilization differ from those obtained for cultivated grasses and legumes. These results provide new insight on the role of seed reserves as energy resources in germination for wild species.

Mobilization and Role of Starch, Protein, and Fat Reserves during Seed Germination of Six Wild Grassland Species

Directory of Open Access Journals (Sweden)

Ming Zhao

2018-02-01

Full Text Available Since seed reserves can influence seed germination, the quantitative and qualitative differences in seed reserves may relate to the germination characteristics of species. The purpose of our study was to evaluate the correlation between germination and seed reserves, as well as their mobilization during germination of six grassland species (Chloris virgata, Kochia scoparia, Lespedeza hedysaroides, Astragalus adsurgens, Leonurus artemisia, and Dracocephalum moldavica and compare the results with domesticated species. We measured starch, protein, and fat content in dry seeds and the initial absorption of water during imbibition. Starch, soluble protein, fat, and soluble sugar content also were determined at five stages during germination. Starch, protein, and fat reserves in dry seeds were not significantly correlated with germination percentage and rate (speed, but soluble sugar and soluble protein contents at different germination stages were positively significantly correlated with germination rate for the six species. Starch was mainly used during seed imbibition, and soluble protein was used from the imbibition stage to the highest germination stage. Fat content for all species remained relatively constant throughout germination for six species, regardless of the proportion of other seed reserves in the seeds. Our results for fat utilization differ from those obtained for cultivated grasses and legumes. These results provide new insight on the role of seed reserves as energy resources in germination for wild species.

Specific leaf mass, fresh: dry weight ratio, sugar and protein contents in species of Lamiaceae from different light environments

Directory of Open Access Journals (Sweden)

M Castrillo

2005-06-01

Full Text Available Samples from eleven species of Lamiaceae were collected from different light environments in Venezuela for laboratory analysis.The studied species were: Plectranthus scutellarioides (Ps, Scutellaria purpurascens (Sp, Hyptis pectinata (Hp, H. sinuata (Hs, Leonorus japonicus (Lj, Plecthranthus amboinicus (Pa Ocimum basilicum (Ocb, O.campechianum (Occ Origanum majorana (Orm, Rosmarinus officinali ,(Ro and Salvia officinalis (So. Protein and soluble sugar contents per unit of area were measured, Specific Leaf Mass (SLMand fresh: dry weight (FW/DW ratios were calculated. The higher values for soluble sugars contents were present in sun species: Lj, Pa, Ocb, Occ, Or. m, Ro and So; the lower values were obtained in low light species: Ps, Sp, Hp, Hs. The values of protein content do not show any clear trend or difference between sun and shade environments. The lowest values for the fresh weight: dry weight ratio are observed in sun species with the exception of Lj and Pa, while the highest value is observed in Pa, a succulent plant. The higher values of specific leaf mass (SLM(Kg DMm-2 are observed in sun plants. The two way ANOVA revealed that there were significant differences among species and between sun and low light environments for sugar content and FW: DW ratio, while SLM was significant for environments but no significant for species, and not significant for protein for both species and environments. The soluble sugar content, FW: DW ratio and SLM values obtained in this work, show a clear separation between sun and shade plants. The sugar content and FW:DW ratio are distinctive within the species,and the light environment affected sugar content, FW:DW ratio and SLM. These species may be shade-tolerant and able to survive in sunny environments. Perhaps these species originated in shaded environments and have been adapting to sunny habitats.Rev.Biol.Trop.53(1-2:23-28.Epub 2005 Jun 24En once especies de la familia Lamiaceae: Plecthranthus

Effective non-denaturing purification method for improving the solubility of recombinant actin-binding proteins produced by bacterial expression.

Science.gov (United States)

Chung, Jeong Min; Lee, Sangmin; Jung, Hyun Suk

2017-05-01

Bacterial expression is commonly used to produce recombinant and truncated mutant eukaryotic proteins. However, heterologous protein expression may render synthesized proteins insoluble. The conventional method used to express a poorly soluble protein, which involves denaturation and refolding, is time-consuming and inefficient. There are several non-denaturing approaches that can increase the solubility of recombinant proteins that include using different bacterial cell strains, altering the time of induction, lowering the incubation temperature, and employing different detergents for purification. In this study, we compared several non-denaturing protocols to express and purify two insoluble 34 kDa actin-bundling protein mutants. The solubility of the mutant proteins was not affected by any of the approaches except for treatment with the detergent sarkosyl. These results indicate that sarkosyl can effectively improve the solubility of insoluble proteins during bacterial expression. Copyright © 2016. Published by Elsevier Inc.

High-Pressure-High-Temperature Processing Reduces Maillard Reaction and Viscosity in Whey Protein-Sugar Solutions

NARCIS (Netherlands)

Avila Ruiz, Geraldine; Xi, Bingyan; Minor, Marcel; Sala, Guido; Boekel, van Tiny; Fogliano, Vincenzo; Stieger, Markus

2016-01-01

The aim of the study was to determine the influence of pressure in high-pressure-high-temperature (HPHT) processing on Maillard reactions and protein aggregation of whey protein-sugar solutions. Solutions of whey protein isolate containing either glucose or trehalose at pH 6, 7, and 9 were

Soluble inhibitors/deactivators of cellulase enzymes from lignocellulosic biomass.

Science.gov (United States)

Kim, Youngmi; Ximenes, Eduardo; Mosier, Nathan S; Ladisch, Michael R

2011-04-07

Liquid hot water, steam explosion, and dilute acid pretreatments of lignocellulose generate soluble inhibitors which hamper enzymatic hydrolysis as well as fermentation of sugars to ethanol. Toxic and inhibitory compounds will vary with pretreatment and include soluble sugars, furan derivatives (hydroxymethyl fulfural, furfural), organic acids (acetic, formic and, levulinic acid), and phenolic compounds. Their effect is seen when an increase in the concentration of pretreated biomass in a hydrolysis slurry results in decreased cellulose conversion, even though the ratio of enzyme to cellulose is kept constant. We used lignin-free cellulose, Solka Floc, combined with mixtures of soluble components released during pretreatment of wood, to prove that the decrease in the rate and extent of cellulose hydrolysis is due to a combination of enzyme inhibition and deactivation. The causative agents were extracted from wood pretreatment liquid using PEG surfactant, activated charcoal or ethyl acetate and then desorbed, recovered, and added back to a mixture of enzyme and cellulose. At enzyme loadings of either 1 or 25mg protein/g glucan, the most inhibitory components, later identified as phenolics, decreased the rate and extent of cellulose hydrolysis by half due to both inhibition and precipitation of the enzymes. Full enzyme activity occurred when the phenols were removed. Hence detoxification of pretreated woods through phenol removal is expected to reduce enzyme loadings, and therefore reduce enzyme costs, for a given level of cellulose conversion. Copyright © 2011 Elsevier Inc. All rights reserved.

Expression and Purification of Soluble STAT5b/STAT3 Proteins for SH2 Domain Binding Assay.

Science.gov (United States)

Asai, Akira; Takakuma, Kazuyuki

2017-01-01

When a large hydrophobic full-length protein is expressed in bacteria, it is often challenging to obtain recombinant proteins in the soluble fraction. One way to overcome this challenge is expression of deletion mutants that have improved solubility while maintaining biological activity. In this chapter, we describe a protocol for expression of truncated forms of STAT5b and STAT3 proteins that are soluble and retain SH2-mediated activity for phospho-Tyr peptide recognition.

Chemical characteristics and enhanced hepatoprotective activities of Maillard reaction products derived from milk protein-sugar system.

Science.gov (United States)

Oh, Nam Su; Young Lee, Ji; Lee, Hyun Ah; Joung, Jae Yeon; Shin, Yong Kook; Kim, Sae Hun; Kim, Younghoon; Lee, Kwang Won

2016-02-01

The objective of this study was to investigate the characteristics, antioxidative properties, and hepatoprotective effects of Maillard reaction products (MRP) from milk protein reacted with sugars. The MRP were obtained from milk protein, whey protein concentrates and sodium caseinate, using 2 types of sugars, lactose and glucose, by heating the mixture at 55°C for 7d in a sodium phosphate buffer (pH 7.4). Changes in the chemical modification of the milk protein were monitored by measuring the protein-bound carbonyls and PAGE protein profiles. The results showed that the amount of protein-bound carbonyls increased after Maillard reaction (MR). In addition, sodium dodecyl sulfate-PAGE analysis indicated a formation of high-molecular weight complexes through MR. The modification sites induced by MR of milk protein were monitored by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry analysis of tryptic-digested gel spots of MRP. As a result, modification and their localization in AA sequence of MRP was identified. Also, the MRP showed higher antioxidant activities than the intact milk protein, and they reduced intracellular reactive oxygen species production and inhibited the depletion of the reduced glutathione concentrations in the HepG2 cells. In particular, glucose-sodium caseinate MRP showed the highest biological activities among all MRP. Therefore, these results suggest that the MRP from milk protein reacting with sugars possess effective antioxidant activity and have a protective ability against oxidative damage. Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Common structural features of cholesterol binding sites in crystallized soluble proteins.

Science.gov (United States)

Bukiya, Anna N; Dopico, Alejandro M

2017-06-01

Cholesterol-protein interactions are essential for the architectural organization of cell membranes and for lipid metabolism. While cholesterol-sensing motifs in transmembrane proteins have been identified, little is known about cholesterol recognition by soluble proteins. We reviewed the structural characteristics of binding sites for cholesterol and cholesterol sulfate from crystallographic structures available in the Protein Data Bank. This analysis unveiled key features of cholesterol-binding sites that are present in either all or the majority of sites: i ) the cholesterol molecule is generally positioned between protein domains that have an organized secondary structure; ii ) the cholesterol hydroxyl/sulfo group is often partnered by Asn, Gln, and/or Tyr, while the hydrophobic part of cholesterol interacts with Leu, Ile, Val, and/or Phe; iii ) cholesterol hydrogen-bonding partners are often found on α-helices, while amino acids that interact with cholesterol's hydrophobic core have a slight preference for β-strands and secondary structure-lacking protein areas; iv ) the steroid's C21 and C26 constitute the "hot spots" most often seen for steroid-protein hydrophobic interactions; v ) common "cold spots" are C8-C10, C13, and C17, at which contacts with the proteins were not detected. Several common features we identified for soluble protein-steroid interaction appear evolutionarily conserved. Copyright © 2017 by the American Society for Biochemistry and Molecular Biology, Inc.

Production of a soluble recombinant prion protein fused to blue fluorescent protein without refolding or detergents in Escherichia coli cells.

Science.gov (United States)

Arii, Yasuhiro; Yamaguchi, Hidenori; Fukuoka, Shin-Ichi

2007-10-01

The physiological function of prion proteins (PrP) remains unclear. To investigate the physiological relevance of PrP, we constructed a fusion protein of PrP with enhanced blue fluorescent protein (PrP-EBFP) to quantify the interaction of PrP with other molecules. Production of soluble PrP-EBFP was achieved by lowering the expression temperature in Escherichia coli (E. coli) cells to 15 degrees C. Soluble PrP-EBFP was purified on cation exchange and heparin-affinity columns to yield high purity protein. This is the first report of the preparation of soluble recombinant PrP without refolding following solubilization using denaturants or disruption using detergents. To confirm the integrity of PrP-EBFP, anisotropy was estimated under physiological conditions in the presence of heparin, which interacts with PrP. The dissociation constant was determined to be 0.88+/-0.07 microM. PrP-EBFP should be useful in the quantification of PrP interactions with other molecules.

Seasonal alteration of sugar metabolism in strawberry ( Fragaria ...

African Journals Online (AJOL)

Plants of strawberry cvs Aromas and Diamante were removed from the field in cold acclimated (CA, January) and non-acclimated (NA, July) stages. Crown parts of the plant were used for analysis. Apoplastic total soluble sugar (TSS), reducing sugars and sucrose contents did not change in both cultivars in both sampling ...

Synthesis of acid-soluble spore proteins by Bacillus subtilis.

OpenAIRE

Leventhal, J M; Chambliss, G H

1982-01-01

The major acid-soluble spore proteins (ASSPs) of Bacillus subtilis were detected by immunoprecipitation of radioactively labeled in vitro- and in vivo-synthesized proteins. ASSP synthesis in vivo began 2 h after the initiation of sporulation (t2) and reached its maximum rate at t7. This corresponded to the time of synthesis of mRNA that stimulated the maximum rate of ASSP synthesis in vitro. Under the set of conditions used in these experiments, protease synthesis began near t0, alkaline phos...

Soluble Protein Analysis using a Compact Bench-top Flow Cytometer

Science.gov (United States)

Pappas, Dimitri; Kao, Shib-Hsin; Cyr, Johnathan

2004-01-01

Future space exploration missions will require analytical technology capable of providing both autonomous medical care to the crew and investigative capabilities to researchers. While several promising candidate technologies exist for further development, flow cytometry is an attractive technology as it offers both crew health (blood cell count, leukocyte differential, etc.) and a wide array of biochemistry and immunology assays. research settings, the application of this technique to soluble protein analysis is also possible. Proteomic beads using fluorescent dyes for optical encoding were used to monitor six cytokines simultaneously in cell medium of cell cultures in stationary and rotating cell culture systems. The results of this work demonstrate that a compact flow cytometer, such as a system proposed for space flight, can detect a variety of soluble proteins for crew health and biotechnology experiments during long-term missions.

A review of machine learning methods to predict the solubility of overexpressed recombinant proteins in Escherichia coli.

Science.gov (United States)

Habibi, Narjeskhatoon; Mohd Hashim, Siti Z; Norouzi, Alireza; Samian, Mohammed Razip

2014-05-08

Over the last 20 years in biotechnology, the production of recombinant proteins has been a crucial bioprocess in both biopharmaceutical and research arena in terms of human health, scientific impact and economic volume. Although logical strategies of genetic engineering have been established, protein overexpression is still an art. In particular, heterologous expression is often hindered by low level of production and frequent fail due to opaque reasons. The problem is accentuated because there is no generic solution available to enhance heterologous overexpression. For a given protein, the extent of its solubility can indicate the quality of its function. Over 30% of synthesized proteins are not soluble. In certain experimental circumstances, including temperature, expression host, etc., protein solubility is a feature eventually defined by its sequence. Until now, numerous methods based on machine learning are proposed to predict the solubility of protein merely from its amino acid sequence. In spite of the 20 years of research on the matter, no comprehensive review is available on the published methods. This paper presents an extensive review of the existing models to predict protein solubility in Escherichia coli recombinant protein overexpression system. The models are investigated and compared regarding the datasets used, features, feature selection methods, machine learning techniques and accuracy of prediction. A discussion on the models is provided at the end. This study aims to investigate extensively the machine learning based methods to predict recombinant protein solubility, so as to offer a general as well as a detailed understanding for researches in the field. Some of the models present acceptable prediction performances and convenient user interfaces. These models can be considered as valuable tools to predict recombinant protein overexpression results before performing real laboratory experiments, thus saving labour, time and cost.

Characterization of soluble dietary fiber from Moringa oleifera seeds and its immunomodulatory effects.

Science.gov (United States)

Anudeep, Sandanamudi; Prasanna, Vaddi K; Adya, Shruthi M; Radha, Cheruppanpullil

2016-10-01

Moringa oleifera (moringa or drumstick) seeds are a potential source of dietary fiber with 6.5% w/w soluble dietary fiber. Biochemical characterization of moringa seed soluble fiber revealed that it is a glycoprotein with 5% neutral sugars. Arabinose and xylose are the major neutral sugars identified by gas liquid chromatography (GLC). Moringa seed soluble fiber was identified as protease resistant-glycoprotein and termed as moringa seed resistant protein (MSRP). MSRP was found to be a homodimer (18kDa) containing two 9kDa monomeric units as revealed by SDS-PAGE analysis with pI 10.8. Immunostimulating activity of MSRP was assessed by murine splenocyte proliferation and production of NO from macrophages. MSRP at low concentration (0.01μg/well) strongly increased proliferation of splenocytes, while MSRP at high concentration weakly responded. MSRP induced 6-fold increase in NO production when compared to the control which indicates the activation of macrophages. MSRP isolated from defatted moringa seed flour is a potent mitogen, enhancing the proliferation of lymphocytes and inducing NO from macrophages. This study concludes that moringa seed is a potential nutritional source to promote the immune system of the host. Copyright © 2016 Elsevier B.V. All rights reserved.

Semen quality and concentration of soluble proteins in the seminal plasma of Alpine bucks Semen quality and concentration of soluble proteins in the seminal plasma of Alpine bucks

Directory of Open Access Journals (Sweden)

Simone Eliza Facione Guimarães

2010-06-01

Full Text Available It was aimed to study the in vitro seminal quality analyzed by complementary tests and to compare them with physical, morphological and biochemical aspects of male goat semen of the Alpine breed. This experiment took place at the Federal University of Viçosa, situated at 20º45’ S latitude and 42º51’ W longitude, Southwest of Brazil. It was done during the summer months of January and February, and three adult male goats of the Alpine breed were used in intensive conditions. The semen was collected by artificial vagina method. In all semen samples (45 ejaculates, after the physical and morphological analysis, the hiposmotic test was done. In 24 ejaculates, it were done thermo-resistance test, and in 21 ejaculates it were determined the concentration of total soluble proteins in seminal plasma. The male goats presented difference in the semen physical and morphological aspects, in the hiposmotic test and thermo-resistance test, but they did not presented difference in total soluble proteins concentration in seminal plasma. Results of the slow thermo-resistance test and hiposmotic test were positively correlated (r = 0.60. It was concluded, according to our results, that the concentration of total soluble proteins in seminal plasma can not be used as a parameter to predict the seminal quality of Alpine bucks.It was aimed to study the in vitro seminal quality analyzed by complementary tests and to compare them with physical, morphological and biochemical aspects of male goat semen of the Alpine breed. This experiment took place at the Federal University of Viçosa, situated at 20º45’ S latitude and 42º51’ W longitude, Southwest of Brazil. It was done during the summer months of January and February, and three adult male goats of the Alpine breed were used in intensive conditions. The semen was collected by artificial vagina method. In all semen samples (45 ejaculates, after the physical and morphological analysis, the hiposmotic test

Salt-soluble proteins from wheat-derived foodstuffs show lower allergenic potency than those from raw flour.

Science.gov (United States)

de Gregorio, Marta; Armentia, Alicia; Díaz-Perales, Araceli; Palacín, Arantxa; Dueñas-Laita, Antonio; Martín, Blanca; Salcedo, Gabriel; Sánchez-Monge, Rosa

2009-04-22

Salt-soluble proteins from wheat flour have been described as main allergens associated with both baker's asthma and food allergy. However, most studies have used raw flour as starting material, thus not considering potential changes in allergenic properties induced by the heat treatment and other industrial processing to produce wheat-derived foodstuffs. Salt extracts from different commercial wheat-derived products were obtained and their allergenic properties investigated by IgE-immunodetection, ELISA assays, and skin prick test. The IgE-binding capacity of salt-soluble proteins from commercial breads and cooked pastas was reduced around 50% compared with that of raw flour, the reduction being less dramatic in noncooked pastas and biscuits. Several wheat-derived foodstuffs showed major IgE-binding components of 20 and 35 kDa, identified as avenin-like and globulin proteins, respectively. These proteins, as well as most flour and bread salt-soluble proteins, were hydrolyzed when subjected to simulated gastrointestinal digestion. However, the digested products still exhibited a residual IgE-binding capacity. Therefore, processing of wheat flour to obtain derived foodstuffs decreases the IgE binding-capacity of the major salt-soluble wheat proteins. Moreover, simulated gastric fluid digestion further inactivates some heat-resistant IgE-binding proteins.

The influence of growth conditions and duckweed diversity on the content of soluble sugar%生长条件和多样性对浮萍可溶性糖含量的影响

Institute of Scientific and Technical Information of China (English)

李阳; 成家杨; 钟钰; 唐杰

2016-01-01

To explore the effects of growth conditions and duckweed species diversity on duckweed growth and soluble sugar productivity,three strains (Lemna aequinoctialis LC33,Landoltia punctata LC06 and Spirodela polyrhiza LC15) were employed to culture either in polyculture or in monocultures under different conditions (temperature,light intensity,nutrient concentration).Results showed that the duckweed accumulated more soluble content and biomass under the low temperature (20 ℃),high light intensity (10 000 lx) and lower nutrient concentration (0 mg N/L,0 mg P/L).Overall,several duckweed polyculture had higher soluble sugar content than those in monoculture.Specifically,the soluble sugar content of the poluculture composed of three strains was the highest when the temperature was low;the soluble sugar content of the polycultures composed of two or three strains was higher than those in monoculture when the light intensity was high;the polyculture composed of two strains accumulated more soluble content under the low nutrient concentration.Therefore,the growth conditions and species diversity had a great influence on soluble sugar content of duckweed.The appropriate polyculture is helpful for improving the biomass production of the whole duckweed cultivation system.%以Lemna aequinoctialis LC33,Landoltia punc tataLC06和Spirodela polyrhiza LC15作为供试材料,在不同环境条件(温度、光照、营养)下对其进行单独和混合培养,研究生长条件和物种多样性对可溶性糖含量和生物量的影响.结果表明,低温(20℃)、强光照(10000lx)、低营养(N为0mg/L,P为0mg/L)的条件更有利于浮萍可溶性糖的积累.此外,适当的混养有助于浮萍可溶性糖的积累,如由3种浮萍组成的混养体系在低温条件下可溶性糖含量较高;由两种或3种浮萍组成的混养体系在强光照条件下可溶性糖含量较高;由两种浮萍组成的混养体系在低营养条件下可溶性糖含量较高.由此可见,生长条件和物种多样性与可溶性糖含量密切相关,适当的混合培养可以提高整个体系的生物质产量.

THE EFFECT OF THE PROTEIN SOLUBILITY OF FISH MEAL AND ...

African Journals Online (AJOL)

utilization of fish meal proteins was determined by a relationship between their solubility or apparent digestibility and the ... Data obtained from these trials should indicate what re- ..... 4 in mind it is thus obvious that the big variation which oc-.

Dynamic analysis of sugar metabolism in different harvest seasons ...

African Journals Online (AJOL)

In pineapple fruits, sugar accumulation plays an important role in flavor characteristics, which varies according to the stage of fruit development. Metabolic changes in the contents of fructose, sucrose and glucose and reducing sugar related to the activities of soluble acid invertase (AI), neutral invertase (NI), sucrose ...

Bromodomain proteins GTE9 and GTE11 are essential for specific BT2-mediated sugar and ABA responses in Arabidopsis thaliana.

Science.gov (United States)

Misra, Anjali; McKnight, Thomas D; Mandadi, Kranthi K

2018-03-01

Global Transcription Factor Group E proteins GTE9 and GTE11 interact with BT2 to mediate ABA and sugar responses in Arabidopsis thaliana. BT2 is a BTB-domain protein that regulates responses to various hormone, stress and metabolic conditions in Arabidopsis thaliana. Loss of BT2 results in plants that are hypersensitive to inhibition of germination by abscisic acid (ABA) and sugars. Conversely, overexpression of BT2 results in resistance to ABA and sugars. Here, we report the roles of BT2-interacting partners GTE9 and GTE11, bromodomain and extraterminal-domain proteins of Global Transcription Factor Group E, in BT2-mediated responses to sugars and hormones. Loss-of-function mutants, gte9-1 and gte11-1, mimicked the bt2-1-null mutant responses; germination of all three mutants was hypersensitive to inhibition by glucose and ABA. Loss of either GTE9 or GTE11 in a BT2 over-expressing line blocked resistance to sugars and ABA, indicating that both GTE9 and GTE11 were required for BT2 function. Co-immunoprecipitation of BT2 and GTE9 suggested that these proteins physically interact in vivo, and presumably function together to mediate responses to ABA and sugar signals.

Activation of human natural killer cells by the soluble form of cellular prion protein

International Nuclear Information System (INIS)

Seong, Yeon-Jae; Sung, Pil Soo; Jang, Young-Soon; Choi, Young Joon; Park, Bum-Chan; Park, Su-Hyung; Park, Young Woo; Shin, Eui-Cheol

2015-01-01

Cellular prion protein (PrP C ) is widely expressed in various cell types, including cells of the immune system. However, the specific roles of PrP C in the immune system have not been clearly elucidated. In the present study, we investigated the effects of a soluble form of recombinant PrP C protein on human natural killer (NK) cells. Recombinant soluble PrP C protein was generated by fusion of human PrP C with the Fc portion of human IgG 1 (PrP C -Fc). PrP C -Fc binds to the surface of human NK cells, particularly to CD56 dim NK cells. PrP C -Fc induced the production of cytokines and chemokines and the degranulation of granzyme B from NK cells. In addition, PrP C -Fc facilitated the IL-15-induced proliferation of NK cells. PrP C -Fc induced phosphorylation of ERK-1/2 and JNK in NK cells, and inhibitors of the ERK or the JNK pathways abrogated PrP C -Fc-induced cytokine production in NK cells. In conclusion, the soluble form of recombinant PrP C -Fc protein activates human NK cells via the ERK and JNK signaling pathways. - Highlights: • Recombinant soluble PrP C (PrP C -Fc) was generated by fusion of human PrP C with IgG1 Fc portion. • PrP C -Fc protein induces the production of cytokines and degranulation from human NK cells. • PrP C -Fc protein enhances the IL-15-induced proliferation of human NK cells. • PrP C -Fc protein activates human NK cells via the ERK and JNK signaling pathways

Variation in total sugars and reductive sugars in the moss Pleurozium schreberi (hylocomiaceae) under water deficit conditions

International Nuclear Information System (INIS)

Montenegro Ruiz, Luis Carlos; Melgarejo Munoz, Luz Marina.

2012-01-01

The structural simplicity of the bryophytes exposed them easily to water stress, forcing them to have physiological and biochemical mechanisms that enable them to survive. This study evaluated the variation of total soluble sugars and reducing sugars in relation to relative water content, in Pleurozium schreberi when faced with low water content in the Paramo de Chingaza (Colombia) and under simulated conditions of water deficit in the laboratory. we found that total sugars increase when the plant is dehydrated and returned to their normal content when re-hydrated moss, this could be interpreted as a possible mechanism of osmotic adjustment and osmoprotection of the cell content and cellular structure. Reducing sugars showed no significant variation, showing that monosaccharides do not have a protective role during dehydration.

A family of E. coli expression vectors for laboratory scale and high throughput soluble protein production

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Bottomley Stephen P

2006-03-01

Full Text Available Abstract Background In the past few years, both automated and manual high-throughput protein expression and purification has become an accessible means to rapidly screen and produce soluble proteins for structural and functional studies. However, many of the commercial vectors encoding different solubility tags require different cloning and purification steps for each vector, considerably slowing down expression screening. We have developed a set of E. coli expression vectors with different solubility tags that allow for parallel cloning from a single PCR product and can be purified using the same protocol. Results The set of E. coli expression vectors, encode for either a hexa-histidine tag or the three most commonly used solubility tags (GST, MBP, NusA and all with an N-terminal hexa-histidine sequence. The result is two-fold: the His-tag facilitates purification by immobilised metal affinity chromatography, whilst the fusion domains act primarily as solubility aids during expression, in addition to providing an optional purification step. We have also incorporated a TEV recognition sequence following the solubility tag domain, which allows for highly specific cleavage (using TEV protease of the fusion protein to yield native protein. These vectors are also designed for ligation-independent cloning and they possess a high-level expressing T7 promoter, which is suitable for auto-induction. To validate our vector system, we have cloned four different genes and also one gene into all four vectors and used small-scale expression and purification techniques. We demonstrate that the vectors are capable of high levels of expression and that efficient screening of new proteins can be readily achieved at the laboratory level. Conclusion The result is a set of four rationally designed vectors, which can be used for streamlined cloning, expression and purification of target proteins in the laboratory and have the potential for being adaptable to a high

Activation of human natural killer cells by the soluble form of cellular prion protein

Energy Technology Data Exchange (ETDEWEB)

Seong, Yeon-Jae [Laboratory of Immunology and Infectious Diseases, Graduate School of Medical Science and Engineering, KAIST, Daejeon (Korea, Republic of); Hafis Clinic, Seoul (Korea, Republic of); Sung, Pil Soo; Jang, Young-Soon; Choi, Young Joon [Laboratory of Immunology and Infectious Diseases, Graduate School of Medical Science and Engineering, KAIST, Daejeon (Korea, Republic of); Park, Bum-Chan [Aging Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon (Korea, Republic of); Park, Su-Hyung [Laboratory of Translational Immunology and Vaccinology, Graduate School of Medical Science and Engineering, KAIST, Daejeon (Korea, Republic of); Park, Young Woo [Aging Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon (Korea, Republic of); Shin, Eui-Cheol, E-mail: ecshin@kaist.ac.kr [Laboratory of Immunology and Infectious Diseases, Graduate School of Medical Science and Engineering, KAIST, Daejeon (Korea, Republic of)

2015-08-21

Cellular prion protein (PrP{sup C}) is widely expressed in various cell types, including cells of the immune system. However, the specific roles of PrP{sup C} in the immune system have not been clearly elucidated. In the present study, we investigated the effects of a soluble form of recombinant PrP{sup C} protein on human natural killer (NK) cells. Recombinant soluble PrP{sup C} protein was generated by fusion of human PrP{sup C} with the Fc portion of human IgG{sub 1} (PrP{sup C}-Fc). PrP{sup C}-Fc binds to the surface of human NK cells, particularly to CD56{sup dim} NK cells. PrP{sup C}-Fc induced the production of cytokines and chemokines and the degranulation of granzyme B from NK cells. In addition, PrP{sup C}-Fc facilitated the IL-15-induced proliferation of NK cells. PrP{sup C}-Fc induced phosphorylation of ERK-1/2 and JNK in NK cells, and inhibitors of the ERK or the JNK pathways abrogated PrP{sup C}-Fc-induced cytokine production in NK cells. In conclusion, the soluble form of recombinant PrP{sup C}-Fc protein activates human NK cells via the ERK and JNK signaling pathways. - Highlights: • Recombinant soluble PrP{sup C} (PrP{sup C}-Fc) was generated by fusion of human PrP{sup C} with IgG1 Fc portion. • PrP{sup C}-Fc protein induces the production of cytokines and degranulation from human NK cells. • PrP{sup C}-Fc protein enhances the IL-15-induced proliferation of human NK cells. • PrP{sup C}-Fc protein activates human NK cells via the ERK and JNK signaling pathways.

Soluble cysteine-rich tick saliva proteins Salp15 and Iric-1 from E. coli.

Science.gov (United States)

Kolb, Philipp; Vorreiter, Jolanta; Habicht, Jüri; Bentrop, Detlef; Wallich, Reinhard; Nassal, Michael

2015-01-01

Ticks transmit numerous pathogens, including borreliae, which cause Lyme disease. Tick saliva contains a complex mix of anti-host defense factors, including the immunosuppressive cysteine-rich secretory glycoprotein Salp15 from Ixodes scapularis ticks and orthologs like Iric-1 from Ixodes ricinus. All tick-borne microbes benefit from the immunosuppression at the tick bite site; in addition, borreliae exploit the binding of Salp15 to their outer surface protein C (OspC) for enhanced transmission. Hence, Salp15 proteins are attractive targets for anti-tick vaccines that also target borreliae. However, recombinant Salp proteins are not accessible in sufficient quantity for either vaccine manufacturing or for structural characterization. As an alternative to low-yield eukaryotic systems, we investigated cytoplasmic expression in Escherichia coli, even though this would not result in glycosylation. His-tagged Salp15 was efficiently expressed but insoluble. Among the various solubility-enhancing protein tags tested, DsbA was superior, yielding milligram amounts of soluble, monomeric Salp15 and Iric-1 fusions. Easily accessible mutants enabled epitope mapping of two monoclonal antibodies that, importantly, cross-react with glycosylated Salp15, and revealed interaction sites with OspC. Free Salp15 and Iric-1 from protease-cleavable fusions, despite limited solubility, allowed the recording of (1)H-(15)N 2D NMR spectra, suggesting partial folding of the wild-type proteins but not of Cys-free variants. Fusion to the NMR-compatible GB1 domain sufficiently enhanced solubility to reveal first secondary structure elements in (13)C/(15)N double-labeled Iric-1. Together, E. coli expression of appropriately fused Salp15 proteins may be highly valuable for the molecular characterization of the function and eventually the 3D structure of these medically relevant tick proteins.

How to find soluble proteins: a comprehensive analysis of alpha/beta hydrolases for recombinant expression in E. coli

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Barth Sandra

2005-04-01

Full Text Available Abstract Background In screening of libraries derived by expression cloning, expression of active proteins in E. coli can be limited by formation of inclusion bodies. In these cases it would be desirable to enrich gene libraries for coding sequences with soluble gene products in E. coli and thus to improve the efficiency of screening. Previously Wilkinson and Harrison showed that solubility can be predicted from amino acid composition (Biotechnology 1991, 9(5:443–448. We have applied this analysis to members of the alpha/beta hydrolase fold family to predict their solubility in E. coli. alpha/beta hydrolases are a highly diverse family with more than 1800 proteins which have been grouped into homologous families and superfamilies. Results The predicted solubility in E. coli depends on hydrolase size, phylogenetic origin of the host organism, the homologous family and the superfamily, to which the hydrolase belongs. In general small hydrolases are predicted to be more soluble than large hydrolases, and eukaryotic hydrolases are predicted to be less soluble in E. coli than prokaryotic ones. However, combining phylogenetic origin and size leads to more complex conclusions. Hydrolases from prokaryotic, fungal and metazoan origin are predicted to be most soluble if they are of small, medium and large size, respectively. We observed large variations of predicted solubility between hydrolases from different homologous families and from different taxa. Conclusion A comprehensive analysis of all alpha/beta hydrolase sequences allows more efficient screenings for new soluble alpha/beta hydrolases by the use of libraries which contain more soluble gene products. Screening of hydrolases from families whose members are hard to express as soluble proteins in E. coli should first be done in coding sequences of organisms from phylogenetic groups with the highest average of predicted solubility for proteins of this family. The tools developed here can be used

Effects of limited irrigation on root yield and quality of sugar beet ...

African Journals Online (AJOL)

STORAGESEVER

2008-12-17

Dec 17, 2008 ... Irrigation treatments had a significant effect on sugar yield and its quality. Potassium ... beet plants respond to water stress by an increase in tap- ..... nitrogen fertilization on soluble carbohydrate concentration in sugar beet.

Brown pigment formation in heated sugar-protein mixed suspensions containing unmodified and peptically modified whey protein concentrates.

Science.gov (United States)

Rongsirikul, Narumol; Hongsprabhas, Parichat

2016-01-01

Commercial whey protein concentrate (WPC) was modified by heating the acidified protein suspensions (pH 2.0) at 80 °C for 30 min and treating with pepsin at 37 °C for 60 min. Prior to spray-drying, such modification did not change the molecular weights (MWs) of whey proteins determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). After spray-drying the modified whey protein concentrate with trehalose excipient (MWPC-TH), it was found that the α-lactalbumin (α-La) was the major protein that was further hydrolyzed the most. The reconstituted MWPC-TH contained β-lactoglobulin (β-Lg) as the major protein and small molecular weight (MW) peptides of less than 6.5 kDa. The reconstituted MWPC-TH had higher NH2 group, Trolox equivalent antioxidant capacity (TEAC), lower exposed aromatic ring and thiol (SH) contents than did the commercial WPC. Kinetic studies revealed that the addition of MWPC-TH in fructose-glycine solution was able to reduce brown pigment formation in the mixtures heated at 80 to 95 °C by increasing the activation energy (Ea) of brown pigment formation due to the retardation of fluoresced advanced glycation end product (AGEs) formation. The addition of MWPC to reducing sugar-glycine/commercial WPC was also able to lower brown pigment formation in the sterilized (121 °C, 15 min) mixed suspensions containing 0.1 M reducing sugar and 0.5-1.0 % glycine and/or commercial (P < 0.05). It was demonstrated that the modification investigated in this study selectively hydrolyzed α-La and retained β-Lg for the production of antibrowning whey protein concentrate.

Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics.

Science.gov (United States)

Hossler, Patrick; McDermott, Sean; Racicot, Christopher; Chumsae, Christopher; Raharimampionona, Haly; Zhou, Yu; Ouellette, David; Matuck, Joseph; Correia, Ivan; Fann, John; Li, Jianmin

2014-01-01

Protein glycosylation is an important post-translational modification toward the structure and function of recombinant therapeutics. The addition of oligosaccharides to recombinant proteins has been shown to greatly influence the overall physiochemical attributes of many proteins. It is for this reason that protein glycosylation is monitored by the developer of a recombinant protein therapeutic, and why protein glycosylation is typically considered a critical quality attribute. In this work, we highlight a systematic study toward the supplementation of sucrose and tagatose into cell culture media for the targeted modulation of protein glycosylation profiles on recombinant proteins. Both sugars were found to affect oligosaccharide maturation resulting in an increase in the percentage of high mannose N-glycan species, as well as a concomitant reduction in fucosylation. The latter effect was demonstrated to increase antibody-dependent cell-mediated cytotoxicity for a recombinant antibody. These aforementioned results were found to be reproducible at different scales, and across different Chinese hamster ovary cell lines. Through the selective supplementation of these described sugars, the targeted modulation of protein glycosylation profiles is demonstrated, as well as yet another tool in the cell culture toolbox for ensuring product comparability. © 2014 American Institute of Chemical Engineers.

Effect of electron radiation on sugar content in inverted liquid sugar

International Nuclear Information System (INIS)

Podadera, P.; Sabato, S.F.

2009-01-01

Inverted liquid sugar is a mixture of sucrose, glucose and fructose, which shows its relevant characteristic on high sweetness power. Ionizing radiation has been applied to different kind of food and ingredients for different reasons, such as pathogens reduction, disinfestations, quarantine purposes, ripening delay among others. Radiation from an electron beam can be utilized as a technique to treat this ingredient because it can process a great volume of material per unit of time. The main goal of this paper was to verify the effect of radiation on the properties of inverted liquid sugar. This ingredient was irradiated in an electron accelerator (Radiation Dynamics) at a dose ranging from 5 to 50 kGy. Sucrose content measurements were reduced by 23% at 30 kGy when compared to control and the reduced sugar content increased around 11%. Density and moisture values were not affected by radiation. The total soluble solids (Brix degrees) rose in function of the absorbed dose. (authors)

BIOACTIVE COMPOUNDS IN CONVENTIONAL AND NO ADDED SUGARS RED STRAWBERRY GUAVA (Psidium cattleianum Sabine JELLIES

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GABRIELA NIEMEYER REISSIG

Full Text Available ABSTRACT This study aimed to prepare jellies of conventional type of red strawberry guava (with added sucrose and no added sugar and evaluate the physical and chemical composition and content of bioactive compounds in them. Four jellies formulations were prepared: conventional with addition of sucrose (F1, aspartame (F2, saccharin and cyclamate (F3, acesulfame and sucralose (F4. Physicochemical analysis of pH were carried out, as well as analysis of titratable acidity, total soluble solids, ashes, proteins, lipids, moisture, carbohydrates, calories, lightness, color tone, total phenols, anthocyanins, carotenoids, ascorbic acid and antioxidant activity, by the capture of DPPH and ABTS radicals. Conventional and no added sugars jellies did not differ for total phenols, total anthocyanins and ascorbic acid. However, processing exerted significant influence (p=0.05 on total carotenoids and antioxidant activity. It is feasible to use red strawberry guava for the preparation of conventional and no added sugar jellies. The products, however, show a significant difference in carotenoids content, with the highest content of these and higher antioxidant activity in processed jellies without sugars addition.

Comparative assessment of sugar and malic acid composition in cultivated and wild apples.

Science.gov (United States)

Ma, Baiquan; Chen, Jie; Zheng, Hongyu; Fang, Ting; Ogutu, Collins; Li, Shaohua; Han, Yuepeng; Wu, Benhong

2015-04-01

Soluble sugar and malic acid contents in mature fruits of 364 apple accessions were quantified using high-performance liquid chromatography (HPLC). Fructose and sucrose represented the major components of soluble sugars in cultivated fruits, whilst fructose and glucose were the major items of sugars in wild fruits. Wild fruits were significantly more acidic than cultivated fruits, whilst the average concentration of total sugars and sweetness index were quite similar between cultivated and wild fruits. Thus, our study suggests that fruit acidity rather than sweetness is likely to have undergone selection during apple domestication. Additionally, malic acid content was positively correlated with glucose content and negatively correlated with sucrose content. This suggests that selection of fruit acidity must have an effect on the proportion of sugar components in apple fruits. Our study provides information that could be helpful for future apple breeding. Copyright © 2014 Elsevier Ltd. All rights reserved.

Fermentable soluble fibres spare amino acids in healthy dogs fed a low-protein diet.

Science.gov (United States)

Wambacq, Wendy; Rybachuk, Galena; Jeusette, Isabelle; Rochus, Kristel; Wuyts, Brigitte; Fievez, Veerle; Nguyen, Patrick; Hesta, Myriam

2016-06-28

Research in cats has shown that increased fermentation-derived propionic acid and its metabolites can be used as alternative substrates for gluconeogenesis, thus sparing amino acids for other purposes. This amino acid sparing effect could be of particular interest in patients with kidney or liver disease, where this could reduce the kidneys'/liver's burden of N-waste removal. Since dogs are known to have a different metabolism than the obligatory carnivorous cat, the main objective of this study was to assess the possibility of altering amino acid metabolism through intestinal fermentation in healthy dogs. This was studied by supplementing a low-protein diet with fermentable fibres, hereby providing an initial model for future studies in dogs suffering from renal/liver disease. Eight healthy dogs were randomly assigned to one of two treatment groups: sugar beet pulp and guar gum mix (SF: soluble fibre, estimated to mainly stimulate propionic acid production) or cellulose (IF: insoluble fibre). Treatments were incorporated into a low-protein (17 %) extruded dry diet in amounts to obtain similar total dietary fibre (TDF) contents for both diets (9.4 % and 8.2 % for the SF and IF diet, respectively) and were tested in a 4-week crossover feeding trial. Apparent faecal nitrogen digestibility and post-prandial fermentation metabolites in faeces and plasma were evaluated. Dogs fed the SF diet showed significantly higher faecal excretion of acetic and propionic acid, resulting in a higher total SCFA excretion compared to IF. SF affected the three to six-hour postprandial plasma acylcarnitine profile by significantly increasing AUC of acetyl-, propionyl-, butyryl- + isobutyryl-, 3-OH-butyryl-, 3-OH-isovaleryl- and malonyl-L-carnitine. Moreover, the amino acid plasma profile at that time was modified as leucine + isoleucine concentrations were significantly increased by SF, and a similar trend for phenylalanine and tyrosine's AUC was found. These results indicate

Combination of sugar analysis and stable isotope ratio mass spectrometry to detect the use of artificial sugars in royal jelly production.

Science.gov (United States)

Wytrychowski, Marine; Daniele, Gaëlle; Casabianca, Hervé

2012-05-01

The effects of feeding bees artificial sugars and/or proteins on the sugar compositions and (13)C isotopic measurements of royal jellies (RJs) were evaluated. The sugars fed to the bees were two C4 sugars (cane sugar and maize hydrolysate), two C3 sugars (sugar beet, cereal starch hydrolysate), and honey. The proteins fed to them were pollen, soybean, and yeast powder proteins. To evaluate the influence of the sugar and/or protein feeding over time, samples were collected during six consecutive harvests. (13)C isotopic ratio measurements of natural RJs gave values of around -25 ‰, which were also seen for RJs obtained when the bees were fed honey or C3 sugars. However, the RJs obtained when the bees were fed cane sugar or corn hydrolysate (regardless of whether they were also fed proteins) gave values of up to -17 ‰. Sugar content analysis revealed that the composition of maltose, maltotriose, sucrose, and erlose varied significantly over time in accordance with the composition of the syrup fed to the bees. When corn and cereal starch hydrolysates were fed to the bees, the maltose and maltotriose contents of the RJs increased up to 5.0 and 1.3 %, respectively, compared to the levels seen in authentic samples (i.e., samples obtained when the bees were fed natural food: honey and pollen) that were inferior to 0.2% and not detected, respectively. The sucrose and erlose contents of natural RJs were around 0.2 %, whereas those in RJs obtained when the bees were fed cane or beet sugar were as much as 4.0 and 1.3 %, respectively. The combination of sugar analysis and (13)C isotopic ratio measurements represents a very efficient analytical methodology for detecting (from early harvests onward) the use of C4 and C3 artificial sugars in the production of RJ.

CCN activation of fumed silica aerosols mixed with soluble pollutants

Science.gov (United States)

Dalirian, M.; Keskinen, H.; Ahlm, L.; Ylisirniö, A.; Romakkaniemi, S.; Laaksonen, A.; Virtanen, A.; Riipinen, I.

2014-09-01

Particle-water interactions of completely soluble or insoluble particles are fairly well understood but less is known of aerosols consisting of mixtures of soluble and insoluble components. In this study, laboratory measurements were performed to investigate cloud condensation nuclei (CCN) activity of silica particles coated with ammonium sulphate (a salt), sucrose (a sugar) and bovine serum albumin known as BSA (a protein). In addition, the agglomerated structure of the silica particles was investigated by estimating the surface equivalent diameter based on measurements with a Differential Mobility Analyzer (DMA) and an Aerosol Particle Mass Analyzer (APM). By using the surface equivalent diameter the non-sphericity of the particles containing silica was accounted for when estimating CCN activation. Furthermore, characterizing critical supersaturations of particles consisting of pure soluble on insoluble compounds using existing frameworks showed that the CCN activation of single component particles was in good agreement with Köhler and adsorption theory based models when the agglomerated structure was accounted for. For mixed particles the CCN activation was governed by the soluble components, and the soluble fraction varied considerably with particle size for our wet-generated aerosols. Our results confirm the hypothesis that knowing the soluble fraction is the key parameter needed for describing the CCN activation of mixed aerosols, and highlight the importance of controlled coating techniques for acquiring a detailed understanding of the CCN activation of atmospheric insoluble particles mixed with soluble pollutants.

Prokaryotic Soluble Overexpression and Purification of Human VEGF165 by Fusion to a Maltose Binding Protein Tag.

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Minh Tan Nguyen

Full Text Available Human vascular endothelial growth factor (VEGF is a key regulator of angiogenesis and plays a central role in the process of tumor growth and metastatic dissemination. Escherichia coli is one of the most common expression systems used for the production of recombinant proteins; however, expression of human VEGF in E. coli has proven difficult because the E. coli-expressed VEGF tends to be misfolded and forms inclusion bodies, resulting in poor solubility. In this study, we successfully produced semi-preparative amounts of soluble bioactive human VEGF165 (hVEGF. We created seven N-terminal fusion tag constructs with hexahistidine (His6, thioredoxin (Trx, glutathione S-transferase (GST, maltose-binding protein (MBP, N-utilization substance protein A (NusA, human protein disulfide isomerase (PDI, and the b'a' domain of PDI (PDIb'a', and tested each construct for soluble overexpression in E. coli. We found that at 18°C, 92.8% of the MBP-tagged hVEGF to be soluble and that this tag significantly increased the protein's solubility. We successfully purified 0.8 mg of pure hVEGF per 500 mL cell culture. The purified hVEGF is stable after tag cleavage, contains very low levels of endotoxin, and is 97.6% pure. Using an Flk1+ mesodermal precursor cell (MPC differentiation assay, we show that the purified hVEGF is not only bioactive but has similar bioactivity to hVEGF produced in mammalian cells. Previous reports on producing hVEGF in E. coli have all been based on refolding of the protein from inclusion bodies. To our knowledge, this is the first report on successfully expressing and purifying soluble hVEGF in E. coli.

Photodamaging mechanism of the eye structure: UV effect on soluble proteins of the lens

International Nuclear Information System (INIS)

Korkhmazyan, M.M.; Fedorovich, I.B.; Ostrovskij, M.A.

1983-01-01

Damaging effect of UV-radiation on soluble proteins of bull lens has been studied. Irradiation results in lens proteins growing yellow, new absorption bands with the maxima 245 and 305 nm appear. It is shown that during photodamage oxidation of SH-groups takes place and protein aggregates are formed

"JCE" Classroom Activity #105. A Sticky Situation: Chewing Gum and Solubility

Science.gov (United States)

Montes-Gonzalez, Ingrid; Cintron-Maldonado, Jose A.; Perez-Medina, Ilia E.; Montes-Berrios, Veronica; Roman-Lopez, Saurie N.

2010-01-01

In this Activity, students perform several solubility tests using common food items such as chocolate, chewing gum, water, sugar, and oil. From their observations during the Activity, students will initially classify the substances tested as soluble or insoluble. They will then use their understanding of the chemistry of solubility to classify the…

Cloning, localization and expression analysis of vacuolar sugar transporters in the CAM plant Ananas comosus (pineapple).

Science.gov (United States)

Antony, Edna; Taybi, Tahar; Courbot, Mikaël; Mugford, Sam T; Smith, J Andrew C; Borland, Anne M

2008-01-01

In photosynthetic tissues of the CAM plant pineapple (Ananas comosus), storage of soluble sugars in the central vacuole during the daytime and their remobilization at night is required to provide carbon skeletons for nocturnal CO(2) fixation. However, soluble sugars produced photosynthetically must also be exported to support growth processes in heterotrophic tissues. To begin to address how vacuolar sugar storage and assimilate partitioning are regulated in A. comosus, degenerate PCR and cDNA library screening were used to clone three candidate sugar transporters from the leaves of this species. Subcellular localization of the three transporters was investigated via expression of YFP-fusion proteins in tobacco epidermal cells and their co-localization with subcellular markers by confocal microscopy. Using this strategy, a putative hexose transporter (AcMST1) and a putative inositol transporter (AcINT1) were identified that both localized to the tonoplast, whereas a putative sucrose transporter (AcSUT1) was found to localize to prevacuolar compartments. A cDNA (AcMST2) with high similarity to a recently characterized tonoplast hexose transporter in Arabidopsis was also identified from an A. comosus fruit EST database. Analyses of transcript abundance indicated that AcMST1 was more highly expressed in fruits compared to leaves of A. comosus, whilst transcripts of AcINT1, AcSUT1, and AcMST2 were more abundant in leaves. Transcript abundance of AcINT1, the putative inositol transporter, showed day-night changes comparable to those of other CAM-related transcripts described in Mesembryanthemum crystallinum. The results are discussed in terms of the role of vacuolar sugar transporters in regulating carbon flow during the diel cycle in CAM plants.

Benzoate-mediated changes on expression profile of soluble proteins in Serratia sp. DS001.

Science.gov (United States)

Pandeeti, E V P; Chinnaboina, M R; Siddavattam, D

2009-05-01

To assess differences in protein expression profile associated with shift in carbon source from succinate to benzoate in Serratia sp. DS001 using a proteomics approach. A basic proteome map was generated for the soluble proteins extracted from Serratia sp. DS001 grown in succinate and benzoate. The differently and differentially expressed proteins were identified using ImageMaster 2D Platinum software (GE Healthcare). The identity of the proteins was determined by employing MS or MS/MS. Important enzymes such as Catechol 1,2 dioxygenase and transcriptional regulators that belong to the LysR superfamily were identified. Nearly 70 proteins were found to be differentially expressed when benzoate was used as carbon source. Based on the protein identity and degradation products generated from benzoate it is found that ortho pathway is operational in Serratia sp. DS001. Expression profile of the soluble proteins associated with shift in carbon source was mapped. The study also elucidates degradation pathway of benzoate in Serratia sp. DS001 by correlating the proteomics data with the catabolites of benzoate.

Anomalous dissolution behaviour of tablets prepared from sugar glass-based solid dispersions

NARCIS (Netherlands)

Van Drooge, D.J.; Hinrichs, W.L.J.; Frijlink, H.W.

2004-01-01

In this study, anomalous dissolution behaviour of tablets consisting of sugar glass dispersions was investigated. The poorly aqueous soluble diazepam was used as a lipophilic model drug. The release of diazepam and sugar carrier was determined to study the mechanisms governing dissolution behaviour.

Properties of spores of Bacillus subtilis strains which lack the major small, acid-soluble protein

International Nuclear Information System (INIS)

Hackett, R.H.; Setlow, P.

1988-01-01

Bacillus subtilis strains containing a deletion in the gene coding for the major small, acid-soluble, spore protein (SASP-gamma) grew and sporulated, and their spores initiated germination normally, but outgrowth of SASP-gamma- spores was significantly slower than that of wild-type spores. The absence of SASP-gamma had no effect on spore protoplast density or spore resistance to heat or radiation. Consequently, SASP-gamma has a different function in spores than do the other major small, acid-soluble proteins

Soluble lens protein polymorphism in flying fishes from the central Arabian Sea

Digital Repository Service at National Institute of Oceanography (India)

Menezes, M.R.

Soluble eye lens nuclei proteins of flying-fishes were studied by cellogel electrophoresis. Three distinct patterns characterized by the number of bands, mobility and staining intensity were observed. Morphological studies of these fishes showed...

Prefoldin Promotes Proteasomal Degradation of Cytosolic Proteins with Missense Mutations by Maintaining Substrate Solubility.

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Sophie A Comyn

2016-07-01

Full Text Available Misfolded proteins challenge the ability of cells to maintain protein homeostasis and can accumulate into toxic protein aggregates. As a consequence, cells have adopted a number of protein quality control pathways to prevent protein aggregation, promote protein folding, and target terminally misfolded proteins for degradation. In this study, we employed a thermosensitive allele of the yeast Guk1 guanylate kinase as a model misfolded protein to investigate degradative protein quality control pathways. We performed a flow cytometry based screen to identify factors that promote proteasomal degradation of proteins misfolded as the result of missense mutations. In addition to the E3 ubiquitin ligase Ubr1, we identified the prefoldin chaperone subunit Gim3 as an important quality control factor. Whereas the absence of GIM3 did not impair proteasomal function or the ubiquitination of the model substrate, it led to the accumulation of the poorly soluble model substrate in cellular inclusions that was accompanied by delayed degradation. We found that Gim3 interacted with the Guk1 mutant allele and propose that prefoldin promotes the degradation of the unstable model substrate by maintaining the solubility of the misfolded protein. We also demonstrated that in addition to the Guk1 mutant, prefoldin can stabilize other misfolded cytosolic proteins containing missense mutations.

Rapid screening for anthocyanins in cane sugars using ESR spectroscopy.

Science.gov (United States)

Thamaphat, Kheamrutai; Goodman, Bernard A; Limsuwan, Pichet; Smith, Siwaporn Meejoo

2015-03-15

Anthocyanin, which is soluble in water and released into sugar steam during extraction, was investigated in this study. The anthocyanin content in refined sugar, plantation white sugar, soft brown sugar and raw sugar was determined using electron spin resonance (ESR) spectroscopy, which was operated at room temperature, and compared with spectra from standard anthocyanin. The ESR spectra of red and violet anthocyanins was predominantly g ≈ 2.0055, which corresponded to an unpaired electron located in the pyrylium ring. Signals for Fe(III) and Mn(II), which naturally occur in plants, were found in raw sugar, soft brown sugar and standard anthocyanin but were absent from refined sugar and plantation white sugar due to the refining process. In addition, the ESR results were correlated with the apparent colour of the sugar, which was determined using the method of the International Commission for Uniform Methods of Sugar Analysis and inductively coupled plasma optical emission spectroscopy. Copyright © 2014 Elsevier Ltd. All rights reserved.

Physicochemical and sensory characteristics of reduced sugar starfruit juice

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Prasetijio, L.D.,

2017-06-01

Full Text Available The objective of this research was to study the physicochemical and sensory characteristics of reduced sugar starfruit juice. Cloudy starfruit juice produced by juicing of the edible part of starfruit, then the obtained filtrate was formulated with citric acid, Na- CMC and different ratio of cane sugar and sorbitol i.e. 10.0%:0.0%, 7.5%:2.5%, 5.0%:5.0%, 2.5%:5.0% and 0.0%:10.0% (% b/v. Physicochemical characteristics of the starfruit juice were total soluble solids in a range of 9.20 and 11.90%, the viscosity of 8.27 and 8.65 cP, pH of 3.56 and 3.63; and titratable acidity of 0.29 and 0.31%. Those cane sugar and sorbitol ratios affected significantly only on the total soluble solids. Based on the sensory evaluation, starfruit juice with cane sugar and sorbitol ratio of 7.50%:2.50% had the highest total preference scores of color, appearance, and taste. Colloidal stability of the starfruit juices during 10 days storage was in a range of 86.26% and 88.18%.

Prokaryotic soluble overexpression and purification of bioactive human growth hormone by fusion to thioredoxin, maltose binding protein, and protein disulfide isomerase.

Directory of Open Access Journals (Sweden)

Minh Tan Nguyen

Full Text Available Human growth hormone (hGH is synthesized by somatotroph cells of the anterior pituitary gland and induces cell proliferation and growth. This protein has been approved for the treatment of various conditions, including hGH deficiency, chronic renal failure, and Turner syndrome. Efficient production of hGH in Escherichia coli (E. coli has proven difficult because the E. coli-expressed hormone tends to aggregate and form inclusion bodies, resulting in poor solubility. In this study, seven N-terminal fusion partners, hexahistidine (His6, thioredoxin (Trx, glutathione S-transferase (GST, maltose-binding protein (MBP, N-utilization substance protein A (NusA, protein disulfide bond isomerase (PDI, and the b'a' domain of PDI (PDIb'a', were tested for soluble overexpression of codon-optimized hGH in E. coli. We found that MBP and hPDI tags significantly increased the solubility of the hormone. In addition, lowering the expression temperature to 18°C also dramatically increased the solubility of all the fusion proteins. We purified hGH from MBP-, PDIb'a'-, or Trx-tagged hGH expressed at 18°C in E. coli using simple chromatographic techniques and compared the final purity, yield, and activity of hGH to assess the impact of each partner protein. Purified hGH was highly pure on silver-stained gel and contained very low levels of endotoxin. On average, ∼37 mg, ∼12 mg, and ∼7 mg of hGH were obtained from 500 mL-cell cultures of Trx-hGH, MBP-hGH, and PDIb'a'-hGH, respectively. Subsequently, hGH was analyzed using mass spectroscopy to confirm the presence of two intra-molecular disulfide bonds. The bioactivity of purified hGHs was demonstrated using Nb2-11 cell.

ESPRIT: an automated, library-based method for mapping and soluble expression of protein domains from challenging targets.

Science.gov (United States)

Yumerefendi, Hayretin; Tarendeau, Franck; Mas, Philippe J; Hart, Darren J

2010-10-01

Expression of sufficient quantities of soluble protein for structural biology and other applications is often a very difficult task, especially when multimilligram quantities are required. In order to improve yield, solubility or crystallisability of a protein, it is common to subclone shorter genetic constructs corresponding to single- or multi-domain fragments. However, it is not always clear where domain boundaries are located, especially when working on novel targets with little or no sequence similarity to other proteins. Several methods have been described employing aspects of directed evolution to the recombinant expression of challenging proteins. These combine the construction of a random library of genetic constructs of a target with a screening or selection process to identify solubly expressing protein fragments. Here we review several datasets from the ESPRIT (Expression of Soluble Proteins by Random Incremental Truncation) technology to provide a view on its capabilities. Firstly, we demonstrate how it functions using the well-characterised NF-kappaB p50 transcription factor as a model system. Secondly, application of ESPRIT to the challenging PB2 subunit of influenza polymerase has led to several novel atomic resolution structures; here we present an overview of the screening phase of that project. Thirdly, analysis of the human kinase TBK1 is presented to show how the ESPRIT technology rapidly addresses the compatibility of challenging targets with the Escherichia coli expression system.

Lack of Detection of Bt Sugarcane Cry1Ab and NptII DNA and Proteins in Sugarcane Processing Products Including Raw Sugar

Directory of Open Access Journals (Sweden)

Adriana Cheavegatti-Gianotto

2018-03-01

Full Text Available Brazil is the largest sugarcane producer and the main sugar exporter in the world. The industrial processes applied by Brazilian mills are very efficient in producing highly purified sugar and ethanol. Literature presents evidence of lack of DNA/protein in these products, regardless of the nature of sugarcane used as raw material. Recently CTNBio, the Brazilian biosafety authority, has approved the first biotechnology-derived sugarcane variety for cultivation, event CTC175-A, which expresses the Cry1Ab protein to control the sugarcane borer (Diatraea saccharalis. The event also expresses neomycin-phosphotransferase type II (NptII protein used as selectable marker during the transformation process. Because of the high purity of sugar and ethanol produced from genetically modified sugarcane, these end-products should potentially be classified as “pure substances, chemically defined,” by Brazilian Biosafety Law No. 11.105. If this classification is to be adopted, these substances are not considered as “GMO derivatives” and fall out of the scope of Law No. 11.105. In order to assess sugar composition and quality, we evaluate Cry1Ab and NptII expression in several sugarcane tissues and in several fractions from laboratory-scale processing of event CTC175-A for the presence of these heterologous proteins as well as for the presence of traces of recombinant DNA. The results of these studies show that CTC175-A presents high expression of Cry1Ab in leaves and barely detectable expression of heterologous proteins in stalks. We also evaluated the presence of ribulose-1,5-bisphosphate carboxylase/oxygenase protein and DNA in the fractions of the industrial processing of conventional Brazilian sugarcane cultivars. Results from both laboratory and industrial processing were concordant, demonstrating that DNA and protein are not detected in the clarified juice and downstream processed fractions, including ethanol and raw sugar, indicating that protein

Synergistic enhancement in the co-gelation of salt-soluble pea proteins and whey proteins.

Science.gov (United States)

Wong, Douglas; Vasanthan, Thava; Ozimek, Lech

2013-12-15

This paper investigated the enhancement of thermal gelation properties when salt-soluble pea proteins were co-gelated with whey proteins in NaCl solutions, using different blend ratios, total protein concentrations, pH, and salt concentrations. Results showed that the thermal co-gelation of pea/whey proteins blended in ratio of 2:8 in NaCl solutions showed synergistic enhancement in storage modulus, gel hardness, paste viscosity and minimum gelation concentrations. The highest synergistic enhancement was observed at pH 6.0 as compared with pH 4.0 and 8.0, and at the lower total protein concentration of 10% as compared with 16% and 22% (w/v), as well as in lower NaCl concentrations of 0.5% and 1.0% as compared with 1.5%, 2.0%, 2.5%, and 3.0% (w/v). The least gelation concentrations were also lower in the different pea/whey protein blend ratios than in pure pea or whey proteins, when dissolved in 1.0% or 2.5% (w/v) NaCl aqueous solutions. Copyright © 2013 Elsevier Ltd. All rights reserved.

Plasma Amino Acid Responses After Consumption of Beverages with Varying Protein Type

Science.gov (United States)

2009-07-01

saturated fat Protein Total earbohydT;ltes dietary fiber soluble fiber total sugar lactose sucrose other carbohydrates (maltodextrin. cocoa powder...provided by the manufacturer, 6 Plasma Amino Acids 7 coagulate for 30 min before being centrifuged for 10 min at 3,600 rpm. Resulting plasma and serum...Therefore, consumers should be aware that these products might not contain the specific protein sources being advertised. Acknowledgment The authors

Enzyme Enzyme activities in relation to sugar accumulation in tomato

International Nuclear Information System (INIS)

Alam, M.J.; Rahman, M.H.; Mamun, M.A.; Islam, K.

2006-01-01

Enzyme activities in tomato juice of five different varieties viz. Ratan, Marglove, BARI-1, BARI-5 and BARI-6, in relation to sugar accumulation were investigated at different maturity stages. The highest amount of invertase and beta-galactosidase was found in Marglove and the lowest in BARI- 6 at all maturity stages. Total soluble sugar and sucrose contents were highest in BARI-1 and lowest in BARI-6. The activity of amylase was maximum in Ratan and minimum in Marglove. Protease activity was highest in Ratan and lowest in BARI-6. BARI-1 contained the highest cellulase activity and the lowest in BARI-5. The amount of total soluble sugar and sucrose increased moderately from premature to ripe stage. The activities of amylase and cellulase increased up to the mature stage and then decreased drastically in the ripe stage. The activities of invertase and protease increased sharply from the premature to the ripe stage while the beta-galactosidase activity decreased remarkably. No detectable amount of reducing sugar was present in the premature stage in all cultivars of tomato but increased thereafter upto the ripe stage. The highest reducing sugar was present in BARI-5 in all of the maturity stages. (author)

Soluble expression of recombinant proteins in the cytoplasm of Escherichia coli

DEFF Research Database (Denmark)

Sørensen, Hans; Mortensen, Kim

2005-01-01

Pure, soluble and functional proteins are of high demand in modern biotechnology. Natural protein sources rarely meet the requirements for quantity, ease of isolation or price and hence recombinant technology is often the method of choice. Recombinant cell factories are constantly employed...... molecular tools available. In spite of all these qualities, expression of recombinant proteins with E. coli as the host often results in insoluble and/or nonfunctional proteins. Here we review new approaches to overcome these obstacles by strategies that focus on either controlled expression of target...... for the production of protein preparations bound for downstream purification and processing. Eschericia coli is a frequently used host, since it facilitates protein expression by its relative simplicity, its inexpensive and fast high density cultivation, the well known genetics and the large number of compatible...

Physicochemical and antioxidant properties of non-refined sugarcane alternatives to white sugar

OpenAIRE

Seguí Gil, Lucía; CALABUIG JIMENEZ, LAURA; Betoret Valls, Noelia; Fito Maupoey, Pedro

2015-01-01

[EN] Antioxidant properties of commercial sugarcane-derived products were analysed to study their suitability for being used as functional ingredients. Cane honey, several jaggeries and several brown sugars were selected from the market and analysed in terms of physicochemical characteristics and antioxidant properties, and compared with white refined sugar (twelve products in total). Moisture, water activity, total soluble solids, pH, colour and sugar profile are reported. As for antioxidant...

Impact of the nutrients N and K and soluble sugars on Diabrotica speciosa (Germar) (Coleoptera, Chrysomelidae) and Agrotis ipsilon (Hufnagel) (Lepidoptera, Noctuidae) populations in potato crops, Solanum tuberosum L. (Solanaceae); Impacto dos nutrientes N e K e de acucares soluveis sobre populacoes de Diabrotica speciosa (Germar) (Coleoptera, Chrysomelidae) e Agrotis ipsilon (Hufnagel) (Lepidoptera, Noctuidae) na cultura da batata, Solanum tuberosum L. (Solanaceae)

Energy Technology Data Exchange (ETDEWEB)

Azeredo, Edson Henrique de [Universidade Federal Fluminense (UFF), Pinheiral, RJ (Brazil). Pro-Reitoria de Extensao], e-mail: edsonhenrique.azeredo@bol.com.br; Lima, Eduardo [Universidade Federal Rural do Rio de Janeiro (UFRRJ), Seropedica, RJ (Brazil). Inst. de Agronomia. Dept. de Solos; Cassino, Paulo Cesar Rodrigues [Universidade Federal Rural do Rio de Janeiro (UFRRJ), Seropedica, RJ (Brazil). Inst. de Biologia. Centro Integrado de Manejo de Pragas C.R.G.

2004-03-15

Impact of the nutrients N and K and soluble sugars on Diabrotica speciosa (Germar) (Coleoptera, Chrysomelidae) and Agrotis ipsilon (Huefnagel) (Lepidoptera, Noctuidae) populations in potato crops, Solanum tuberosum L. (Solanaceae). The occurrence of Diabrotica speciosa (Germar, 1824) and Agrotis ipsilon (Huefnagel, 1767) on the potato cultivars Achat and Monalisa, influenced by nitrogen and potassium dosage, and minimum quantity of soluble sugars, was studied. The following parameters were evaluated: concentration of mineral nutrient and sugar in green leaf, senescent leaf, leaf in abscission, stem, tubercle and total plant using extracts of infusion in ethanol 80%. The largest infestation of D. speciosa larvae was on Monalisa cultivar at 150 kg.ha{sup -1} of N + K with 27.03% at P< 0,05. It was observed that the effect of the dosage of N + K in the increment of the concentration of soluble sugars increased the damages in the tubercles and stems by A. ipsilon. The infestation by these species increased to 58.82% on the Monalisa cultivar, when the nitrogen dosage increased from zero to 150 kg.ha{sup -1}, in the absence of potassium. On the other hand, high dosage of K reduced the damages by A. ipsilon on Monalisa cultivar. However, it did not influence the storage of soluble sugar. The results indicated that in Achat cultivar the accumulated soluble sugar was reduced, probably sensitized by elevation of potassic fertilization dosing, differing from Monalisa cultivar, in which the influence was by nitrogen dosing. (author)

An autoclave treatment reduces the solubility and antigenicity of an allergenic protein found in buckwheat flour.

Science.gov (United States)

Tomotake, Hiroyuki; Yamazaki, Rikio; Yamato, Masayuki

2012-06-01

The effects of an autoclave treatment of buckwheat flour on a 24-kDa allergenic protein were investigated by measuring reduction in solubility and antibody binding. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed that the intensity of the major bands, including that of the 24-kDa allergen, was reduced by the autoclave treatment. The protein solubility in buckwheat flour was variably decreased by the autoclave treatment. Enzyme-linked immunosorbent assay analysis using a monoclonal antibody specific for buckwheat 24-kDa protein showed that the reactivity of protein extracts (10 μg/ml) from buckwheat flour was lowered by the autoclave treatment. The autoclave treatment may reduce the major allergen content of buckwheat. Future studies will determine if autoclaving treatments affect the allergenicity of the 24-kDa buckwheat protein.

Sugar recognition by human galactokinase

Directory of Open Access Journals (Sweden)

Timson David J

2003-11-01

Full Text Available Abstract Background Galactokinase catalyses the first committed step of galactose catabolism in which the sugar is phosphorylated at the expense of MgATP. Recent structural studies suggest that the enzyme makes several contacts with galactose – five side chain and two main chain hydrogen bonds. Furthermore, it has been suggested that inhibition of galactokinase may help sufferers of the genetic disease classical galactosemia which is caused by defects in another enzyme of the pathway galactose-1-phosphate uridyl transferase. Galactokinases from different sources have a range of substrate specificities and a diversity of kinetic mechanisms. Therefore only studies on the human enzyme are likely to be of value in the design of therapeutically useful inhibitors. Results Using recombinant human galactokinase expressed in and purified from E. coli we have investigated the sugar specificity of the enzyme and the kinetic consequences of mutating residues in the sugar-binding site in order to improve our understanding of substrate recognition by this enzyme. D-galactose and 2-deoxy-D-galactose are substrates for the enzyme, but N-acetyl-D-galactosamine, L-arabinose, D-fucose and D-glucose are all not phosphorylated. Mutation of glutamate-43 (which forms a hydrogen bond to the hydroxyl group attached to carbon 6 of galactose to alanine results in only minor changes in the kinetic parameters of the enzyme. Mutation of this residue to glycine causes a ten-fold drop in the turnover number. In contrast, mutation of histidine 44 to either alanine or isoleucine results in insoluble protein following expression in E. coli. Alteration of the residue that makes hydrogen bonds to the hydroxyl attached to carbons 3 and 4 (aspartate 46 results in an enzyme that although soluble is essentially inactive. Conclusions The enzyme is tolerant to small changes at position 2 of the sugar ring, but not at positions 4 and 6. The results from site directed mutagenesis could

Transport of soluble proteins through the Golgi occurs by diffusion via continuities across cisternae

Science.gov (United States)

Beznoussenko, Galina V; Parashuraman, Seetharaman; Rizzo, Riccardo; Polishchuk, Roman; Martella, Oliviano; Di Giandomenico, Daniele; Fusella, Aurora; Spaar, Alexander; Sallese, Michele; Capestrano, Maria Grazia; Pavelka, Margit; Vos, Matthijn R; Rikers, Yuri GM; Helms, Volkhard; Mironov, Alexandre A; Luini, Alberto

2014-01-01

The mechanism of transport through the Golgi complex is not completely understood, insofar as no single transport mechanism appears to account for all of the observations. Here, we compare the transport of soluble secretory proteins (albumin and α1-antitrypsin) with that of supramolecular cargoes (e.g., procollagen) that are proposed to traverse the Golgi by compartment progression–maturation. We show that these soluble proteins traverse the Golgi much faster than procollagen while moving through the same stack. Moreover, we present kinetic and morphological observations that indicate that albumin transport occurs by diffusion via intercisternal continuities. These data provide evidence for a transport mechanism that applies to a major class of secretory proteins and indicate the co-existence of multiple intra-Golgi trafficking modes. DOI: http://dx.doi.org/10.7554/eLife.02009.001 PMID:24867214

Dynamics of sugar-metabolic enzymes and sugars accumulation during watermelon (citrullus lanatus) fruit development

International Nuclear Information System (INIS)

Zhang, H.

2016-01-01

We analyzed sugar accumulation and the activities of sugar-metabolic enzymes in ripening fruits of three cultivars of watermelon; a high-sugar type w2, a low-sugar type (w1), and their hybrid. In w2, the glucose and fructose contents were higher than the sucrose content in the earlier stage of fruit development, and fruit growth was accompanied by increases in glucose, fructose, and sucrose contents. The sucrose content increased substantially after 20 days after anthesis (DAA) and it was the main soluble sugar in mature fruit (sucrose: hexoses ratio, 0.71). In W, the fructose and glucose contents were significantly higher than the sucrose content in mature fruit (sucrose: hexoses ratio, 0.25). Comparing the two parent cultivars, sucrose was the most important factor affecting the total sugar content in mature fruit, although glucose and fructose also contributed to total sugar contents. The fructose and glucose contents in the fruit of F1 were mid-way between those of their parents, while the sucrose content was closer to that of W (sucrose:hexoses ratio in F1, 0.26). In the early stage of fruit development of W2, the activities of acid invertase and neutral invertase were higher than those of sucrose synthase and sucrose phosphate synthase. After 20 DAA, the acid invertase and neutral invertase activities decreased and those of sucrose synthase and sucrose phosphate synthase increased, leading to increased sucrose content. In W1, the activities of acid invertase and neutral invertase were higher than those of sucrose synthase and sucrose phosphate synthase at the early stage. The sucrose synthase and sucrose phosphate synthase activities were lower in W1 than in W2 at the later stages of fruit development. The patterns of sugar accumulation and sugar-metabolic enzyme activities during fruit development in F1 were similar to those in W1. (author)

High-throughput method for optimum solubility screening for homogeneity and crystallization of proteins

Science.gov (United States)

Kim, Sung-Hou [Moraga, CA; Kim, Rosalind [Moraga, CA; Jancarik, Jamila [Walnut Creek, CA

2012-01-31

An optimum solubility screen in which a panel of buffers and many additives are provided in order to obtain the most homogeneous and monodisperse protein condition for protein crystallization. The present methods are useful for proteins that aggregate and cannot be concentrated prior to setting up crystallization screens. A high-throughput method using the hanging-drop method and vapor diffusion equilibrium and a panel of twenty-four buffers is further provided. Using the present methods, 14 poorly behaving proteins have been screened, resulting in 11 of the proteins having highly improved dynamic light scattering results allowing concentration of the proteins, and 9 were crystallized.

Effects of protein versus simple sugar intake on weight loss in polycystic ovary syndrome (according to the National Institutes of Health criteria).

Science.gov (United States)

Kasim-Karakas, Sidika E; Almario, Rogelio U; Cunningham, Wendy

2009-07-01

To compare the effects of protein vs. simple sugars on weight loss, body composition, and metabolic and endocrine parameters in polycystic ovary syndrome (PCOS). A 2-month, free-living, randomized, single-blinded study. University PCOS clinic. Thirty-three patients with PCOS. To achieve a final energy reduction of 450 kcal/day, first the daily energy intake was reduced by 700 kcal; then a 240-kcal supplement containing either whey protein or simple sugars was added. Changes in weight, fat mass, fasting glucose and insulin, plasma lipoproteins, and sex steroids. Twenty-four subjects (13 in the simple sugars group and 11 in the protein group) completed the study. The protein group lost more weight (-3.3 +/- 0.8 kg vs. -1.1 +/- 0.6 kg) and more fat mass (-3.1 +/- 0.9 kg vs. -0.5 +/- 0.6 kg) and had larger decreases in serum cholesterol (-33.0 +/- 8.4 mg/dL vs. -2.3 +/- 6.8 mg/dL), high-density lipoprotein cholesterol (-4.5 +/- 1.3 mg/dL vs. -0.4 +/- 1.3 mg/dL), and apoprotein B (-20 +/- 5 mg/dL vs. 3 +/- 5 mg/dL). In patients with PCOS, a hypocaloric diet supplemented with protein reduced body weight, fat mass, serum cholesterol, and apoprotein B more than the diet supplemented with simple sugars.

Arabidopsis scaffold protein RACK1A modulates rare sugar D-allose regulated gibberellin signaling.

Science.gov (United States)

Fennell, Herman; Olawin, Abdulquadri; Mizanur, Rahman M; Izumori, Ken; Chen, Jin-Gui; Ullah, Hemayet

2012-11-01

As energy sources and structural components, sugars are the central regulators of plant growth and development. In addition to the abundant natural sugars in plants, more than 50 different kinds of rare sugars exist in nature, several of which show distinct roles in plant growth and development. Recently, one of the rare sugars, D-allose, an epimer of D-glucose at C3, is found to suppress plant hormone gibberellin (GA) signaling in rice. Scaffold protein RACK1A in the model plant Arabidopsis is implicated in the GA pathway as rack1a knockout mutants show insensitivity to GA in GA-induced seed germination. Using genetic knockout lines and a reporter gene, the functional role of RACK1A in the D-allose pathway was investigated. It was found that the rack1a knockout seeds showed hypersensitivity to D-allose-induced inhibition of seed germination, implicating a role for RACK1A in the D-allose mediated suppression of seed germination. On the other hand, a functional RACK1A in the background of the double knockout mutations in the other two RACK1 isoforms, rack1b/rack1c, showed significant resistance to the D-allose induced inhibition of seed germination. The collective results implicate the RACK1A in the D-allose mediated seed germination inhibition pathway. Elucidation of the rare sugar signaling mechanism will help to advance understanding of this less studied but important cellular signaling pathway.

Effects of Watering and Fertilization on Carbohydrate Reserves in Sugar Maple Seedlings

Science.gov (United States)

Clayton M., Jr. Carl; John R. Donnelly; Boyd W. Post

1978-01-01

Sugar maple seedlings, grown under three nutrient and three moisture levels, were analyzed after three growing seasons for starch and ethanol-soluble sugars. Analytical procedures are detailed in the appendix. Fertilization did not affect carbohydrate levels in stems or roots. Water stress caused a significant reduction in the amount of carbohydrates in stems and roots...

In meso in situ serial X-ray crystallography of soluble and membrane proteins

International Nuclear Information System (INIS)

Huang, Chia-Ying; Olieric, Vincent; Ma, Pikyee; Panepucci, Ezequiel; Diederichs, Kay; Wang, Meitian; Caffrey, Martin

2015-01-01

A method for performing high-throughput in situ serial X-ray crystallography with soluble and membrane proteins in the lipid cubic phase is described. It works with microgram quantities of protein and lipid (and ligand when present) and is compatible with the most demanding sulfur SAD phasing. The lipid cubic phase (LCP) continues to grow in popularity as a medium in which to generate crystals of membrane (and soluble) proteins for high-resolution X-ray crystallographic structure determination. To date, the PDB includes 227 records attributed to the LCP or in meso method. Among the listings are some of the highest profile membrane proteins, including the β 2 -adrenoreceptor–G s protein complex that figured in the award of the 2012 Nobel Prize in Chemistry to Lefkowitz and Kobilka. The most successful in meso protocol to date uses glass sandwich crystallization plates. Despite their many advantages, glass plates are challenging to harvest crystals from. However, performing in situ X-ray diffraction measurements with these plates is not practical. Here, an alternative approach is described that provides many of the advantages of glass plates and is compatible with high-throughput in situ measurements. The novel in meso in situ serial crystallography (IMISX) method introduced here has been demonstrated with AlgE and PepT (alginate and peptide transporters, respectively) as model integral membrane proteins and with lysozyme as a test soluble protein. Structures were solved by molecular replacement and by experimental phasing using bromine SAD and native sulfur SAD methods to resolutions ranging from 1.8 to 2.8 Å using single-digit microgram quantities of protein. That sulfur SAD phasing worked is testament to the exceptional quality of the IMISX diffraction data. The IMISX method is compatible with readily available, inexpensive materials and equipment, is simple to implement and is compatible with high-throughput in situ serial data collection at macromolecular

Evidence that cellulolysis by an anaerobic ruminal fungus is catabolite regulated by glucose, cellobiose, and soluble starch

International Nuclear Information System (INIS)

Morrison, M.; Mackie, R.I.; Kistner, A.

1990-01-01

A Piromyces-like ruminal fungus was used to study preferential carbohydrate utilization of [U- 14 C]cellulose, both alone and in combination with several soluble sugars. For cells grown on cellulose alone, cellulolytic activity was immediate and, initially, greater than that observed in the presence of added carbohydrate. Cellulolytic activity remained minimal in cultures containing cellulose plus glucose or cellobiose until the soluble sugar was depleted. Soluble starch also regulated cellulose activity but to a lesser extent. The results presented suggest that some fungal cellulases are susceptible to catabolite regulatory mechanisms

Effects of Toasting Time on Digestive Hydrolysis of Soluble and Insoluble 00-Rapeseed Meal Proteins

NARCIS (Netherlands)

Salazar-Villanea, Sergio; Bruininx, Erik M.A.M.; Gruppen, Harry; Carré, Patrick; Quinsac, Alain; Poel, van der Thomas

2017-01-01

Thermal damage to proteins can reduce their nutritional value. The effects of toasting time on the kinetics of hydrolysis, the resulting molecular weight distribution of 00-rapeseed meal (RSM) and the soluble and insoluble protein fractions separated from the RSM were studied. Hydrolysis was

Rapid Determination of Protein Solubility and Stability Conditions for NMR Studies Using Incomplete Factorial Design

International Nuclear Information System (INIS)

Ducat, Thierry; Declerck, Nathalie; Gostan, Thierry; Kochoyan, Michel; Demene, Helene

2006-01-01

Sample preparation constitutes a crucial and limiting step in structural studies of proteins by NMR. The determination of the solubility and stability (SAS) conditions of biomolecules at millimolar concentrations stays today empirical and hence time- and material-consuming. Only few studies have been recently done in this field and they have highlighted the interest of using crystallogenesis tools to optimise sample conditions. In this study, we have adapted a method based on incomplete factorial design and making use of crystallisation plates to quantify the influence of physico-chemical parameters such as buffer pH and salts on protein SAS. A description of the experimental set up and an evaluation of the method are given by case studies on two functional domains from the bacterial regulatory protein LicT as well as two other proteins. Using this method, we could rapidly determine optimised conditions for extracting soluble proteins from bacterial cells and for preparing purified protein samples sufficiently concentrated and stable for NMR characterisation. The drastic reduction in the time and number of experiments required for searching protein SAS conditions makes this method particularly well-adapted for a systematic investigation on a large range of physico-chemical parameters

Sugar-rich sweet sorghum is distinctively affected by wall polymer features for biomass digestibility and ethanol fermentation in bagasse.

Science.gov (United States)

Li, Meng; Feng, Shengqiu; Wu, Leiming; Li, Ying; Fan, Chunfen; Zhang, Rui; Zou, Weihua; Tu, Yuanyuan; Jing, Hai-Chun; Li, Shizhong; Peng, Liangcai

2014-09-01

Sweet sorghum has been regarded as a typical species for rich soluble-sugar and high lignocellulose residues, but their effects on biomass digestibility remain unclear. In this study, we examined total 63 representative sweet sorghum accessions that displayed a varied sugar level at stalk and diverse cell wall composition at bagasse. Correlative analysis showed that both soluble-sugar and dry-bagasse could not significantly affect lignocellulose saccharification under chemical pretreatments. Comparative analyses of five typical pairs of samples indicated that DP of crystalline cellulose and arabinose substitution degree of non-KOH-extractable hemicelluloses distinctively affected lignocellulose crystallinity for high biomass digestibility. By comparison, lignin could not alter lignocellulose crystallinity, but the KOH-extractable G-monomer predominately determined lignin negative impacts on biomass digestions, and the G-levels released from pretreatments significantly inhibited yeast fermentation. The results also suggested potential genetic approaches for enhancing soluble-sugar level and lignocellulose digestibility and reducing ethanol conversion inhibition in sweet sorghum. Copyright © 2014. Published by Elsevier Ltd.

Glycan Reader is improved to recognize most sugar types and chemical modifications in the Protein Data Bank.

Science.gov (United States)

Park, Sang-Jun; Lee, Jumin; Patel, Dhilon S; Ma, Hongjing; Lee, Hui Sun; Jo, Sunhwan; Im, Wonpil

2017-10-01

Glycans play a central role in many essential biological processes. Glycan Reader was originally developed to simplify the reading of Protein Data Bank (PDB) files containing glycans through the automatic detection and annotation of sugars and glycosidic linkages between sugar units and to proteins, all based on atomic coordinates and connectivity information. Carbohydrates can have various chemical modifications at different positions, making their chemical space much diverse. Unfortunately, current PDB files do not provide exact annotations for most carbohydrate derivatives and more than 50% of PDB glycan chains have at least one carbohydrate derivative that could not be correctly recognized by the original Glycan Reader. Glycan Reader has been improved and now identifies most sugar types and chemical modifications (including various glycolipids) in the PDB, and both PDB and PDBx/mmCIF formats are supported. CHARMM-GUI Glycan Reader is updated to generate the simulation system and input of various glycoconjugates with most sugar types and chemical modifications. It also offers a new functionality to edit the glycan structures through addition/deletion/modification of glycosylation types, sugar types, chemical modifications, glycosidic linkages, and anomeric states. The simulation system and input files can be used for CHARMM, NAMD, GROMACS, AMBER, GENESIS, LAMMPS, Desmond, OpenMM, and CHARMM/OpenMM. Glycan Fragment Database in GlycanStructure.Org is also updated to provide an intuitive glycan sequence search tool for complex glycan structures with various chemical modifications in the PDB. http://www.charmm-gui.org/input/glycan and http://www.glycanstructure.org. wonpil@lehigh.edu. Supplementary data are available at Bioinformatics online. © The Author (2017). Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com

Soluble expression of recomb inant cMyc, Klf4, Oct4, and Sox2 proteins in bacteria and transduction into living cells

Directory of Open Access Journals (Sweden)

Guo-Dan Liu

2017-04-01

Full Text Available AIM: To develop a new method to produce recombinant reprogramming proteins, cMyc, Klf4, Oct4, and Sox2, in soluble format with low cost for the generation of induced pluripotent stem cells (iPSCs. METHODS: A short polypeptide sequence derived from the HIV trans-activator of transcription protein (TAT and the nucleus localization signal (NLS polypeptide were fused to the N terminus of the reprogramming proteins and they were constructed into pCold-SUMO vector which can extremely improve the solubility of recombinant proteins. Then these vector plasmids were transformed into E. coli BL21 (DE3 Chaperone competent cells for amplification. The solubility of these recombinant proteins was determined by SDS-PAGE and Coomassie brilliant blue staining. The recombinant proteins were purified by Ni-NTA resin and identified by Western blot. The transduction of these proteins into HEK 293T cells were evaluated by immunofluorescence staining. RESULTS: These four reprogramming proteins could be produced in soluble format in pCold-SUMO expression vector system with the assistance of chaperone proteins in bacteria. The proteins were purified successfully with a purity of over 70% with a relative high transduction rate into 293 cells. CONCLUSION: The results in the present study indicate the four important reprogramming proteins, cMyc, Klf4, Oct4, and Sox2, can be produced in soluble format in bacteria with low cost. Our new method thus might be expected to greatly contribute to the future study of iPSCs.

Amides are novel protein modifications formed by physiological sugars.

Science.gov (United States)

Glomb, M A; Pfahler, C

2001-11-09

The Maillard reaction, or nonenzymatic browning, proceeds in vivo, and the resulting protein modifications (advanced glycation end products) have been associated with various pathologies. Despite intensive research only very few structures have been established in vivo. We report here for the first time N(6)-[2-[(5-amino-5-carboxypentyl)amino]-2-oxoethyl]lysine (GOLA) and N(6)-glycoloyllysine (GALA) as prototypes for novel amide protein modifications produced by reducing sugars. Their identity was confirmed by independent synthesis and coupled liquid chromatography/mass spectrometry. Model reactions with N(alpha)-t-butoxycarbonyl-lysine showed that glyoxal and glycolaldehyde are immediate precursors, and reaction pathways are directly linked to N(epsilon)-carboxymethyllysine via glyoxal-imine structures. GOLA, the amide cross-link, and 1,3-bis(5-amino-5-carboxypentyl)imidazolium salt (GOLD), the imidazolium cross-link, share a common intermediate. The ratio of GOLA to GOLD is greater when glyoxal levels are low at constant lysine concentrations. GOLA and GALA formation from the Amadori product of glucose and lysine depends directly upon oxidation. With the advanced glycation end product inhibitors aminoguanidine and pyridoxamine we were able to dissect oxidative fragmentation of the Amadori product as a second mechanism of GOLA formation exactly coinciding with N(epsilon)-carboxymethyllysine synthesis. In contrast, the formation of GALA appears to depend solely upon glyoxal-imines. After enzymatic hydrolysis GOLA was found at 66 pmol/mg of brunescent lens protein. This suggests amide protein modifications as important markers of pathophysiological processes.

Ambient aerosol concentrations of sugars and sugar-alcohols at four different sites in Norway

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K. E. Yttri

2007-08-01

Full Text Available Sugars and sugar-alcohols are demonstrated to be important constituents of the ambient aerosol water-soluble organic carbon fraction, and to be tracers for primary biological aerosol particles (PBAP. In the present study, levels of four sugars (fructose, glucose, sucrose, trehalose and three sugar-alcohols (arabitol, inositol, mannitol in ambient aerosols have been quantified using a novel HPLC/HRMS-TOF (High Performance Liquid Chromatography in combination with High Resolution Mass Spectrometry – Time of Flight method to assess the contribution of PBAP to PM>sub>10 and PM_2.5. Samples were collected at four sites in Norway at different times of the year in order to reflect the various contributing sources and the spatial and seasonal variation of the selected compounds.

Sugars and sugar-alcohols were present at all sites investigated, underlining the ubiquity of these highly polar organic compounds. The highest concentrations were reported for sucrose, reaching a maximum concentration of 320 ng m⁻³ in PM₁₀ and 55 ng m⁻³ in PM_2.5. The mean concentration of sucrose was up to 10 times higher than fructose, glucose and the dimeric sugar trehalose. The mean concentrations of the sugar-alcohols were typically lower, or equal, to that of the monomeric sugars and trehalose. Peak concentrations of arabitol and mannitol did not exceed 30 ng m⁻³ in PM₁₀, and for PM_2.5 all concentrations were below 6 ng m⁻³.

Sugars and sugar-alcohols were associated primarily with coarse aerosols except during wintertime at the suburban site in Elverum, where a shift towards sub micron aerosols was observed. It is proposed that this shift was due to the intensive use of wood burning for residential heating at this site during winter, confirmed by high concurrent concentrations of levoglucosan. Elevated concentrations of sugars in PM₂

Performance of dairy goats fed diets with dry yeast from sugar cane as protein source

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Luciano Soares de Lima

2012-01-01

Full Text Available The effects of inactive dry yeast (Saccharomyces cerevisiae from sugar cane were studied in 18 primiparus Saanen dairy goats (51.07±1.43 on dry matter intake and digestibility, milk production and quality. Animals were distributed in a completely randomized design during 90 days (from day 60 of milking. Diets were composed of soybean meal; soybean meal + dry yeast; or dry yeast, as protein sources, and ground corn, mineral supplement and corn silage (40%. Animals fed the dry yeast diet showed lower intake of dry matter (DM, organic matter (OM, crude protein, ether extract and neutral detergent fiber. Diets did not influence milk yield; however the milk production efficiency (kg of milk produced/kg of crude protein ingested was better in goats fed the dry yeast diet. Acidity, somatic cell counts and milk urea nitrogen values were not affected by treatments. Animals fed the soybean + dry yeast diet had higher fat and total solids than those fed the dry yeast diet. The digestibility of DM, OM and total carbohydrate was lower for soybean only and soybean + dry yeast diets. Total digestible nutrients were higher for dry yeast and soy bean diets than soybean + dry yeast diet. Dry yeast from sugar cane is a good alternative protein source for feeding lactating dairy goats and can be recommended because it maintains the production performance.

Elastin binds to a multifunctional 67-kilodalton peripheral membrane protein

International Nuclear Information System (INIS)

Mecham, R.P.; Hinek, A.; Entwistle, R.; Wrenn, D.S.; Griffin, G.L.; Senior, R.M.

1989-01-01

Elastin binding proteins from plasma membranes of elastin-producing cells were isolated by affinity chromatography on immobilized elastin peptides. Three proteins of 67, 61, and 55 kDa were released from the elastin resin by guanidine/detergent, soluble elastin peptides, synthetic peptide VGVAPG, or galactoside sugars, but not by synthetic RGD-containing peptide or sugars not related to galactose. All three proteins incorporated radiolabel upon extracellular iodination and contained [ 3 H]leucine following metabolic labeling, confirming that each is a synthetic product of the cell. The 67-kDa protein could be released from the cell surface with lactose-containing buffers, whereas solubilization of the 61- and 55-kDa components required the presence of detergent. Although all three proteins were retained on elastin affinity columns, the 61- and 55-kDa components were retained only in the presence of 67-kDa protein, suggesting that the 67-kDa protein binds elastin and the 61- and 55-kDa proteins bind to the 67-kDa protein. The authors propose that the 67-, 61-, and 55-kDa proteins constitute an elastin-receptor complex that forms a transmembrane link between the extracellular matrix and the intracellular compartment

Improving the process of progressive preliming in sugar production

OpenAIRE

V. A. Golybin; V. A. Fedoruk; N. A. Voronkova; I. S. Bushmin

2016-01-01

Tasks progressive preliming are to conduct neutralization, coagulation, deposition of non-sugars, the formation of a precipitate structure. This is the first operation of physical and chemical cleaning of the raw juice in the case of it in optimum conditions enables the coagulation and transferred to precipitate a substantial portion of high-molecular non-sugars, poorly soluble calcium salts that provides the cleaning effect of 14–18%, that is, up to half of the total effect on the whole stat...

Antioxidative activity of water soluble polysaccharide in pumpkin fruits (Cucurbita maxima Duchesne).

Science.gov (United States)

Nara, Kazuhiro; Yamaguchi, Akira; Maeda, Naomi; Koga, Hidenori

2009-06-01

We evaluated the antioxidative activity of a water soluble polysaccharide fraction (WSP) from pumpkin fruits (Cucurbita maxima Duchesne). In the WSP, DPPH radical scavenging and superoxide dismutase-like activity increased depending on the total sugar content. Furthermore, the WSP can serve as an inhibitor of ascorbic acid oxidation. The efficacy was also affected by the total sugar content.

Molecular dynamics simulations of lysozyme in water/sugar solutions

Energy Technology Data Exchange (ETDEWEB)

Lerbret, A. [Department of Food Science, Cornell University, 101 Stocking Hall, Ithaca, NY 14853 (United States); Affouard, F. [Laboratoire de Dynamique et Structure des Materiaux Moleculaires, UMR CNRS 8024, Universite Lille I, 59655 Villeneuve d' Ascq Cedex (France)], E-mail: frederic.affouard@univ-lille1.fr; Bordat, P. [Laboratoire de Chimie Theorique et de Physico-Chimie Moleculaire, UMR 5624, Universite de Pau et des Pays de l' Adour, 64000 Pau (France); Hedoux, A.; Guinet, Y.; Descamps, M. [Laboratoire de Dynamique et Structure des Materiaux Moleculaires, UMR CNRS 8024, Universite Lille I, 59655 Villeneuve d' Ascq Cedex (France)

2008-04-18

Structural and dynamical properties of the solvent at the protein/solvent interface have been investigated by molecular dynamics simulations of lysozyme in trehalose, maltose and sucrose solutions. Results are discussed in the framework of the bioprotection phenomena. The analysis of the relative concentration of water oxygen atoms around lysozyme suggests that lysozyme is preferentially hydrated. When comparing the three sugars, trehalose is seen more excluded than maltose and sucrose. The preferential exclusion of sugars from the protein surface induces some differences in the behavior of trehalose and maltose, particularly at 50 and 60 wt% concentrations, that are not observed experimentally in binary sugar/mixtures. The dynamical slowing down of the solvent is suggested to mainly arise from the homogeneity of the water/sugar matrices controlled by the percolation of the sugar hydrogen bonds networks. Furthermore, lysozyme strongly increases relaxation times of solvent molecules at the protein/solvent interface.

Effect of plaster cast immobilization on the turnover rates of soluble proteins and lactate dehydrogenase isoenzymes of rabbit M. soleus

Energy Technology Data Exchange (ETDEWEB)

Edes, I.; Dosa, E.; Sohar, I.; Guba, F. (Orvostudomanyi Egyetem, Szeged (Hungary). Biokemiai Tanszek)

1982-01-01

In atrophized muscle the decreases of the activity of LDH isoenzymes can be explained partly by a 15 per cent decrease of the enzyme synthesis and partly by a 25 per cent increase in catabolism. The quantities of the soluble proteins and LDH were measured after intravenously administered /sup 3/H-leucin incorporation, from the musculus soleus. LDH was isolated by means of affinity chromatography. Radioactivity was determined in a Packard Tri-Carb scintillation counter. The synthesis rate of soluble proteins barely changed during immobilization. In the atrophized muscle the decrease of the amount of soluble proteins could be almost exclusively interpreted in terms of a 25 per cent enchancement of degradative process. The accelerated catabolism is most probably due to the proteolytic enzymes activated by immobilization.

The effect of honey compared to sucrose, mixed sugars, and a sugar-free diet on weight gain in young rats.

Science.gov (United States)

Chepulis, L M

2007-04-01

To determine whether honey, sucrose, and mixed sugars as in honey have different effects on weight gain, 40 6-wk-old Sprague-Dawley rats were fed a powdered diet that was either sugar free or contained 8% sucrose, 8% mixed sugars as in honey, or 10% honey freely for 6 wk. Weight gain and food intake were assessed weekly, and at completion of the study blood samples were removed for measurement of blood sugar (HbA1c) and a fasting lipid profile. The animals were then minced and total percentage body fat and protein measured. Overall percentage weight gain was significantly lower in honey-fed rats than those fed sucrose or mixed sugars, despite a similar food intake. Weight gains were comparable for rats fed honey and a sugar free diet although food intake was significantly higher in honey-fed rats. HbA1c and triglyceride levels were significantly higher in all sugar treatments compared with rats fed a sugar free diet, but no other differences in lipid profiles were reported. No differences in percentage body fat or protein levels were reported.

Co-Production of Fungal Biomass Derived Constituents and Ethanol from Citrus Wastes Free Sugars without Auxiliary Nutrients in Airlift Bioreactor.

Science.gov (United States)

Satari, Behzad; Karimi, Keikhosro; Taherzadeh, Mohammad J; Zamani, Akram

2016-02-26

The potential of two zygomycetes fungi, Mucor indicus and Rhizopus oryzae, in assimilating citrus waste free sugars (CWFS) and producing fungal chitosan, oil, and protein as well as ethanol was investigated. Extraction of free sugars from citrus waste can reduce its environmental impact by decreasing the possibility of wild microorganisms growth and formation of bad odors, a typical problem facing the citrus industries. A total sugar concentration of 25.1 g/L was obtained by water extraction of citrus waste at room temperature, used for fungal cultivation in shake flasks and airlift bioreactor with no additional nutrients. In shake flasks cultivations, the fungi were only able to assimilate glucose, while fructose remained almost intact. In contrast, the cultivation of M. indicus and R. oryzae in the four-liter airlift bioreactor resulted in the consumption of almost all sugars and production of 250 and 280 g fungal biomass per kg of consumed sugar, respectively. These biomasses correspondingly contained 40% and 51% protein and 9.8% and 4.4% oil. Furthermore, the fungal cell walls, obtained after removing the alkali soluble fraction of the fungi, contained 0.61 and 0.69 g chitin and chitosan per g of cell wall for M. indicus and R. oryzae, respectively. Moreover, the maximum ethanol yield of 36% and 18% was obtained from M. indicus and R. oryzae, respectively. Furthermore, that M. indicus grew as clump mycelia in the airlift bioreactor, while R. oryzae formed spherical suspended pellets, is a promising feature towards industrialization of the process.

Co-Production of Fungal Biomass Derived Constituents and Ethanol from Citrus Wastes Free Sugars without Auxiliary Nutrients in Airlift Bioreactor

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Behzad Satari

2016-02-01

Full Text Available The potential of two zygomycetes fungi, Mucor indicus and Rhizopus oryzae, in assimilating citrus waste free sugars (CWFS and producing fungal chitosan, oil, and protein as well as ethanol was investigated. Extraction of free sugars from citrus waste can reduce its environmental impact by decreasing the possibility of wild microorganisms growth and formation of bad odors, a typical problem facing the citrus industries. A total sugar concentration of 25.1 g/L was obtained by water extraction of citrus waste at room temperature, used for fungal cultivation in shake flasks and airlift bioreactor with no additional nutrients. In shake flasks cultivations, the fungi were only able to assimilate glucose, while fructose remained almost intact. In contrast, the cultivation of M. indicus and R. oryzae in the four-liter airlift bioreactor resulted in the consumption of almost all sugars and production of 250 and 280 g fungal biomass per kg of consumed sugar, respectively. These biomasses correspondingly contained 40% and 51% protein and 9.8% and 4.4% oil. Furthermore, the fungal cell walls, obtained after removing the alkali soluble fraction of the fungi, contained 0.61 and 0.69 g chitin and chitosan per g of cell wall for M. indicus and R. oryzae, respectively. Moreover, the maximum ethanol yield of 36% and 18% was obtained from M. indicus and R. oryzae, respectively. Furthermore, that M. indicus grew as clump mycelia in the airlift bioreactor, while R. oryzae formed spherical suspended pellets, is a promising feature towards industrialization of the process.

Cleaved thioredoxin fusion protein enables the crystallization of poorly soluble ERα in complex with synthetic ligands

International Nuclear Information System (INIS)

Cura, Vincent; Gangloff, Monique; Eiler, Sylvia; Moras, Dino; Ruff, Marc

2007-01-01

A new crystallization strategy: the presence of cleaved thioredoxin fusion is critical for crystallization of the estrogen nuclear receptor ligand binding domain in complex with synthetic ligands. This novel technique should be regarded as an interesting alternative for crystallization of difficult proteins. The ligand-binding domain (LBD) of human oestrogen receptor α was produced in Escherichia coli as a cleavable thioredoxin (Trx) fusion in order to improve solubility. Crystallization trials with either cleaved and purified LBD or with the purified fusion protein both failed to produce crystals. In another attempt, Trx was not removed from the LBD after endoproteolytic cleavage and its presence promoted nucleation and subsequent crystal growth, which allowed the structure determination of two different LBD–ligand–coactivator peptide complexes at 2.3 Å resolution. This technique is likely to be applicable to other low-solubility proteins

1181-IJBCS-Article-Gandonou G Christophe

African Journals Online (AJOL)

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the world. In this study, the effects of salt stress on free proline, soluble sugars and soluble proteins ..... Table 1: Results of 2-ways analysis of variance of proline and soluble sugars accumulation in leaves and ..... 1; Release 6.03). Edn. SAS.

Flux, rejection and fouling during microfiltration and ultrafiltration of sugar palm sap using a pilot plant scale

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Wanichapichart, P.

2006-07-01

Full Text Available The possibility of using a pilot plant scale microfiltration (MF and ultrafiltration (UF to clarify and reduce number of bacteria, yeast and mould of sugar palm sap was studied. The membrane used was multi channel tubular ceramic membrane (ZrO2-TiO2 with membrane pore size 0.2 and 0.1 μm and molecular weight cut off (MWCO 300 and 50 kDa for microfiltration and ultrafiltration respectively. The experiment was carried out to investigate the rejection of the components in sugar palm sap, permeate flux and fouling characteristics. The results showed that the turbidity, the total solid, the viscosity and the numbers of bacteria, yeast and mould in the permeate obtained by MF and UF were reduced significantly compared to those of fresh sugar palm sap. The total soluble solid, total sugar, reducing sugar and pH were not affected by MF and UF. The permeate fluxes for all membranes were reduced greatly as the volume concentration ratio (VCR increased due to severe fouling. The irreversible fouling on membrane surface and/or inside the membrane tended to increase with increasing membrane pore size or MWCO. The result also suggested that protein and small particle in the sugar palm sap were probably responsible for the internal fouling of large pore size membrane. According to the physical, chemical and microorganism quality results, both MF and UF showed the potential use for improving the quality of sugar palm sap but flux reduction due to fouling was a major problem affecting the process performance.

Increasing the Thermostable Sugar-1-Phosphate Nucleotidylyltransferase Activities of the Archaeal ST0452 Protein through Site Saturation Mutagenesis of the 97th Amino Acid Position.

Science.gov (United States)

Honda, Yuki; Zang, Qian; Shimizu, Yasuhiro; Dadashipour, Mohammad; Zhang, Zilian; Kawarabayasi, Yutaka

2017-02-01

The ST0452 protein is a bifunctional protein exhibiting sugar-1-phosphate nucleotidylyltransferase (sugar-1-P NTase) and amino-sugar-1-phosphate acetyltransferase activities and was isolated from the thermophilic archaeon Sulfolobus tokodaii Based on the previous observation that five single mutations increased ST0452 sugar-1-P NTase activity, nine double-mutant ST0452 proteins were generated with the intent of obtaining enzymes exhibiting a further increase in catalysis, but all showed less than 15% of the wild-type N-acetyl-d-glucosamine-1-phosphate uridyltransferase (GlcNAc-1-P UTase) activity. The Y97A mutant exhibited the highest activity of the single-mutant proteins, and thus site saturation mutagenesis of the 97th position (Tyr) was conducted. Six mutants showed both increased GlcNAc-1-P UTase and glucose-1-phosphate uridyltransferase activities, eight mutants showed only enhanced GlcNAc-1-P UTase activity, and six exhibited higher GlcNAc-1-P UTase activity than that of the Y97A mutant. Kinetic analyses of three typical mutants indicated that the increase in sugar-1-P NTase activity was mainly due to an increase in the apparent k cat value. We hypothesized that changing the 97th position (Tyr) to a smaller amino acid with similar electronic properties would increase activity, and thus the Tyr at the corresponding 103rd position of the Escherichia coli GlmU (EcGlmU) enzyme was replaced with the same residues. The Y103N mutant EcGlmU showed increased GlcNAc-1-P UTase activity, revealing that the Tyr at the 97th position of the ST0452 protein (103rd position in EcGlmU) plays an important role in catalysis. The present results provide useful information regarding how to improve the activity of natural enzymes and how to generate powerful enzymes for the industrial production of sugar nucleotides. It is typically difficult to increase enzymatic activity by introducing substitutions into a natural enzyme. However, it was previously found that the ST0452 protein

Synthesis of acid-soluble spore proteins by Bacillus subtilis.

Science.gov (United States)

Leventhal, J M; Chambliss, G H

1982-12-01

The major acid-soluble spore proteins (ASSPs) of Bacillus subtilis were detected by immunoprecipitation of radioactively labeled in vitro- and in vivo-synthesized proteins. ASSP synthesis in vivo began 2 h after the initiation of sporulation (t2) and reached its maximum rate at t7. This corresponded to the time of synthesis of mRNA that stimulated the maximum rate of ASSP synthesis in vitro. Under the set of conditions used in these experiments, protease synthesis began near t0, alkaline phosphatase synthesis began at about t2, and refractile spores were first observed between t7 and t8. In vivo- and in vitro-synthesized ASSPs comigrated in sodium dodecyl sulfate-polyacrylamide gels. Their molecular weights were 4,600 (alpha and beta) and 11,000 (gamma). The average half-life of the ASSP messages was 11 min when either rifampin (10 micrograms/ml) or actinomycin D (1 microgram/ml) was used to inhibit RNA synthesis.

An electrophoretic study of the soluble lens proteins from the Indian mackerel, Rastrelliger kanagurta (Cuv)

Digital Repository Service at National Institute of Oceanography (India)

Menezes, M.R.

Soluble eye lens nuclei proteins of the Indian mackerel Rastrelliger kanagurta were studied by cellogel electrophoresis, to see whether there are any intra species variations. A distinct pattern characterised by the number of bands, mobility...

Protein shedding in urothelial bladder cancer: prognostic implications of soluble urinary EGFR and EpCAM.

Science.gov (United States)

Bryan, R T; Regan, H L; Pirrie, S J; Devall, A J; Cheng, K K; Zeegers, M P; James, N D; Knowles, M A; Ward, D G

2015-03-17

Better biomarkers must be found to develop clinically useful urine tests for bladder cancer. Proteomics can be used to identify the proteins released by cancer cell lines and generate candidate markers for developing such tests. We used shotgun proteomics to identify proteins released into culture media by eight bladder cancer cell lines. These data were compared with protein expression data from the Human Protein Atlas. Epidermal growth factor receptor (EGFR) was identified as a candidate biomarker and measured by ELISA in urine from 60 noncancer control subjects and from 436 patients with bladder cancer and long-term clinical follow-up. Bladder cancer cell lines shed soluble EGFR ectodomain. Soluble EGFR is also detectable in urine and is highly elevated in some patients with high-grade bladder cancer. Urinary EGFR is an independent indicator of poor bladder cancer-specific survival with a hazard ratio of 2.89 (95% CI 1.81-4.62, Pbladder cancer-specific survival and have prognostic value over and above that provided by standard clinical observations. Measuring urinary EGFR and EpCAM may represent a simple and useful approach for fast-tracking the investigation and treatment of patients with the most aggressive bladder cancers.

Sugar beet leaves: from biorefinery to techno-functionality

NARCIS (Netherlands)

Kiskini, Alexandra

2017-01-01

Sugar beet leaves (SBL), which are a side stream of the sugar beets cultivation, are currently left unexploited after sugar beets have been harvested. The general aim of this thesis was to study the biorefinery of SBL, with a special focus on the isolation of proteins. To reach this aim the

Heat-Induced Soluble Protein Aggregates from Mixed Pea Globulins and β-Lactoglobulin.

Science.gov (United States)

Chihi, Mohamed-Lazhar; Mession, Jean-luc; Sok, Nicolas; Saurel, Rémi

2016-04-06

The present work investigates the formation of protein aggregates (85 °C, 60 min incubation) upon heat treatment of β-lactoglobulin (βlg)-pea globulins (Glob) mixtures at pH 7.2 and 5 mM NaCl from laboratory-prepared protein isolates. Various βlg/Glob weight ratios were applied, for a total protein concentration of 2 wt % in admixture. Different analytical methods were used to determine the aggregation behavior of "mixed" aggregates, that is, surface hydrophobicity and also sulfhydryl content, protein interactions by means of SDS-PAGE electrophoresis, and molecule size distribution by DLS and gel filtration. The production of "mixed" thermal aggregates would involve both the formation of new disulfide bonds and noncovalent interactions between the denatured βlg and Glob subunits. The majority of "mixed" soluble aggregates displayed higher molecular weight and smaller diameter than those for Glob heated in isolation. The development of pea-whey protein "mixed" aggregates may help to design new ingredients for the control of innovative food textures.

Water-Soluble Dried Blood Spot in Protein Analysis: A Proof-of-Concept Study.

Science.gov (United States)

Rosting, Cecilie; Gjelstad, Astrid; Halvorsen, Trine Grønhaug

2015-08-04

In the present work human chorionic gonadotropin (hCG) was used as a model protein in a proof-of-concept study combining water-soluble dried blood spot (DBS) material in liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based protein analysis. A water-soluble material consisting of commercially available carboxymethyl cellulose (CMC) was evaluated as sampling material for this purpose. The material dissolved readily at physiological pH. Different sample preparation methods were evaluated, and in the final method, 15 μL of whole blood was deposited and dried on CMC before the whole spot was dissolved prior to cleanup by immunoaffinity extraction, tryptic digest, and preconcentration by solid-phase extraction (SPE). The results indicated complete dissolution of hCG from the spots, acceptable limit of detection (LOD) (0.1 IU/mL), linearity (R(2) = 0.959), accuracy (16%), and precision (≤22%). Long-term stability (45 days) of hCG in dried spots at reduced temperatures (≤8 °C) was also demonstrated. The analyte recovery was comparable to the commercially available nonsolvable cellulose material (FTA DMPK-C card).

Soluble arabinoxylan alters digesta flow and protein digestion of red meat-containing diets in pigs.

Science.gov (United States)

Zhang, Dagong; Williams, Barbara A; Mikkelsen, Deirdre; Li, Xiuhua; Keates, Helen L; Lisle, Allan T; Collins, Helen M; Fincher, Geoffrey B; Bird, Anthony R; Topping, David L; Gidley, Michael J; Bryden, Wayne L

2015-09-01

The aim of this study was to investigate how a moderate increase in dietary meat content combined (or not) with soluble fibre would influence protein digestion as well as digesta characteristics and flow. Four groups of pigs were fed Western-style diets (high-protein/high-fat) containing two types of barbecued red meat, one with and one without a wheat arabinoxylan-rich fraction. After 4 wk, digesta samples were collected from small and large intestinal sites and analyzed for protein, amino acids, dry matter, and acid-insoluble ash. Tissue samples were also collected from each site. Arabinoxylan consumption led to somewhat lower apparent protein digestibility within the small and large intestines as well as shorter mean retention times. This suggests that the lowered protein digestibility is due, at least partly, to shorter access time to digestive proteases and absorptive surfaces. Additionally, digesta mass was higher in pigs fed arabinoxylan while dry matter (%) was lower, indicating an increased digesta water-holding capacity due to the presence of a soluble dietary fiber. Data showed that solubilized wheat arabinoxylan provides potential health benefits through decreased protein digestibility, increased digesta mass, and reduced mean retention time, even for diets with a moderately higher protein content. These factors are associated with efficiency of digestion and satiety, both of which have implications for prevention of obesity and other health disorders. Copyright © 2015 Elsevier Inc. All rights reserved.

Alcohol from sugar beets

Energy Technology Data Exchange (ETDEWEB)

Malchenko, A L; Verzhbitskaya, V A

1962-01-01

The factor which determines the economy in the EtOH industry which uses sugar beets as raw materials is the rapid and complete recovery of the sugar contained in the beets for fermentation purposes. It is best to extract the beets at 70 to 75/sup 0/. Thorough shredding of the beets then need no longer form part of the operation, and the protein compounds, which give rise to fuel oils, are extracted in small amounts only.

THE TECHNOLOGY OF PASTE FROM SUGAR BEET OBTAINING

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M. G. Magomedov

2014-01-01

Full Text Available We considered a new technology for processing sugar beet into intermediate product for the food industry. Nowadays sugar beet is mainly used for processing into granulated sugar. In the granulated sugar obtaining sugar beet undergoes deep extraction of all nutrients: proteins, minerals, pectin, organic acids, preventing the crystallization process, and then the refined product (granulated sugar with a mass fraction of sucrose not less than 99.75% is obtained. We developed a technology for producing a sugar beet paste, which allows to preserve both sucrose, and almost all useful for human food substances containing in beet. A sugar beet paste is a valuable food product. Carbohydrates, organic acids, minerals, proteins, dietary fibers and vitamins are found in it. The block diagram of a sugar beet paste is given in the article. Technology of obtaining a sugar beet paste was tested under production conditions at the cannery (Joint Stock Company "Sadovoye" Liskinsky district,. Storozhevoe village, Voronezh region. The chemical composition of the paste (Dry Substances = 40% was determined, the degree of meeting of daily requirement for nutrients was calculated. 100 g of the product satisfy the daily requirement for dietary fiber by 42.5%, organic acids by 27.5% potassium by 24.0%, magnesium by 40.0%, iron by 26.7%. The sugar beet paste can be used as a semi-finished product in the confectionery, bakery, food concentrates industry, for products with increased nutritional value, as well as the finished product instead of marmalade and jam.

Utilization of paramagnetic relaxation enhancements for high-resolution NMR structure determination of a soluble loop-rich protein with sparse NOE distance restraints

International Nuclear Information System (INIS)

Furuita, Kyoko; Kataoka, Saori; Sugiki, Toshihiko; Hattori, Yoshikazu; Kobayashi, Naohiro; Ikegami, Takahisa; Shiozaki, Kazuhiro; Fujiwara, Toshimichi; Kojima, Chojiro

2015-01-01

NMR structure determination of soluble proteins depends in large part on distance restraints derived from NOE. In this study, we examined the impact of paramagnetic relaxation enhancement (PRE)-derived distance restraints on protein structure determination. A high-resolution structure of the loop-rich soluble protein Sin1 could not be determined by conventional NOE-based procedures due to an insufficient number of NOE restraints. By using the 867 PRE-derived distance restraints obtained from the NOE-based structure determination procedure, a high-resolution structure of Sin1 could be successfully determined. The convergence and accuracy of the determined structure were improved by increasing the number of PRE-derived distance restraints. This study demonstrates that PRE-derived distance restraints are useful in the determination of a high-resolution structure of a soluble protein when the number of NOE constraints is insufficient

Protein expression in the midgut of sugar-fed Aedes albopictus females

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Saboia-Vahia Leonardo

2012-12-01

Full Text Available Abstract Background Aedes albopictus is a vector for several fatal arboviruses in tropical and sub-tropical regions of the world. The midgut of the mosquito is the first barrier that pathogens must overcome to establish infection and represents one of the main immunologically active sites of the insect. Nevertheless, little is known about the proteins involved in the defense against pathogens, and even in the processing of food, and the detoxification of metabolites. The identification of proteins exclusively expressed in the midgut is the first step in understanding the complex physiology of this tissue and can provide insight into the mechanisms of pathogen-vector interaction. However, identification of the locally expressed proteins presents a challenge because the Ae. albopictus genome has not been sequenced. Methods In this study, two-dimensional electrophoresis (2DE was combined with liquid chromatography in line with tandem mass spectrometry (LC-MS/MS and data mining to identify the major proteins in the midgut of sugar-fed Ae. albopictus females. Results Fifty-six proteins were identified by sequence similarity to entries from the Ae. aegypti genome. In addition, two hypothetical proteins were experimentally confirmed. According to the gene ontology analysis, the identified proteins were classified into 16 clusters of biological processes. Use of the STRING database to investigate protein functional associations revealed five functional networks among the identified proteins, including a network for carbohydrate and amino acid metabolism, a group associated with ATP production and a network of proteins that interact during detoxification of toxic free radicals, among others. This analysis allowed the assignment of a potential role for proteins with unknown function based on their functional association with other characterized proteins. Conclusion Our findings represent the first proteome map of the Ae. albopictus midgut and denotes the

Soluble proteins from fowl feather keratin. II. Isolation of some proteins from barbs.

Science.gov (United States)

Murayama, K; Akahane, K; Murozono, S

1977-01-01

Four fractions, GF-1, 2, 3, and 4, which had been separated from S-carboxymethylated (SCM-) proteins of fowl feathers by gel filtration, were each chromatographed on a DEAE-cellulose column in 0.05 M Tris-HCl buffer (pH 8.5) containing 8 M urea. The major fraction, GF-3, was further separated into seven peaks; the first four were shown to be single components by polyacrylamide disc gel electrophoresis. Chromatograms of GF-1 and 2 showed broad peaks which appeared at nearly the same volume as in GF-3. The components from GF-3 had very similar amino acid compositions except that the SCM-cysteine content showed a tendency to increase in the order of elution from the column. SCM-extract prepared from barbs of the wing feathers of a fowl was more heterogeneous than that taken from the body feathers. A combination of gel filtration on Sephadex G-75 and chromatography on DEAE-cellulose was found to be more effective for the isolation of soluble SCM-proteins.

A novel water-based process produces eco-friendly bio-adhesive made from green cross-linked soybean soluble polysaccharide and soy protein.

Science.gov (United States)

Yuan, Cheng; Chen, Mingsong; Luo, Jing; Li, Xiaona; Gao, Qiang; Li, Jianzhang

2017-08-01

In this study, an eco-friendly soy protein adhesive was developed that utilized two components from soybean meal without addition of any toxic material. A plant-based, water-soluble and inexpensive soybean soluble polysaccharide was used as the novel renewable material to combine with soy protein to produce a soy protein adhesive. Three-plywood was fabricated with the resulting adhesive, and its wet shear strength was measured. The results showed the wet shear strength of plywood bonded by the adhesive reached 0.99MPa, meeting the water resistance requirement for interior use panels. This improvement was attributed to the following reasons: (1) Combination of cross-linked soybean soluble polysaccharide and soy protein formed an interpenetrating network structure, improving the thermal stability and water resistance of the cured adhesive. (2) Adding CL-SSPS decreased the adhesive viscosity to 15.14Pas, which increased the amount of the adhesive that penetrate the wood's surface and formed more interlocks. Copyright © 2017 Elsevier Ltd. All rights reserved.

Production of ethanol in batch and fed-batch fermentation of soluble sugar

International Nuclear Information System (INIS)

Chaudhary, M.Y.; Shah, M.A.; Shah, F.H.

1991-01-01

Keeping in view of the demand and need for alternate energy source, especially liquid fuels and the availability of raw materials in Pakistan, we have carried out biochemical and technological studies for ethanol through fermentation of renewable substrates. Molasses and sugar cane have been used as substrate for yeast fermentation. Selected yeast were used in both batch and semi continuous fermentation of molasses. Clarified dilute molasses were fermented with different strains of Saccharomyces cerevisiae. Ethanol concentration after 64 hours batch fermentation reached 9.4% with 90% yield based on sugar content. During feed batch system similar results were obtained after a fermentation cycle of 48 hours resulting in higher productivity. Similarly carbohydrates in fruit juices and hydro lysates of biomass can be economically fermented to ethanol to be used as feed stock for other chemicals. (author)

Spray Drying of High Sugar Content Foods: Improving of Product Yield and Powder Properties

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Mehmet Koç

2016-05-01

Full Text Available Spray drying is the most preferred drying method to produce powdered food in the food industry and it is also widely used to convert sugar-rich liquid foods to a powder form. During and/or after spray drying process of sugar-rich products, undesirable situation was appeared such as stickiness, high moisture affinity (hygroscopicity and low solubility due to low molecular weight monosaccharides that found naturally in the structure. The basis of these problems was formed by low glass transition temperature of sugar-rich products. This review gives information about the difficulties in drying of sugar-rich products via spray dryer, actions need to be taken against these difficulties and drying of sugar-rich honey and fruit juices with spray drying method.

Characterization of the Water-Soluble Fraction of Woody Biomass Pyrolysis Oils

Energy Technology Data Exchange (ETDEWEB)

Stankovikj, Filip; McDonald, Armando G.; Helms, Gregory L.; Olarte, Mariefel V.; Garcia-Perez, Manuel

2017-01-31

This paper reports a study of the chemical composition of the water soluble (WS) fraction obtained by cold water precipitation of two commercial wood pyrolysis oils (BTG and Amaron). The fraction studied accounts for between 50.3 and 51.3 wt. % of the oils. With the most common analytical techniques used today for the characterization of this fraction (KF titration, GC/MS, hydrolysable sugars and total carbohydrates), it is possible to quantify only between 45 and 50 wt. % of it. Our results confirm that most of the total carbohydrates (hydrolysable sugars and non-hydrolysable) are soluble in water. The ion chromatography hydrolysis method showed that between 11.6 and 17.3 wt. % of these oils were hydrolysable sugars. A small quantity of phenols detectable by GC/MS (between 2.5 and 3.9 wt. %) were identified. It is postulated that the unknown high molecular weight fraction (30-55 wt. %) is formed by highly dehydrated sugars rich in carbonyl groups and WS phenols. The overall content of carbonyl, carboxyl, hydroxyl and phenolic compounds in the WS fraction were quantified by titration, Folin-Ciocalteu, 31P-NMR and 1H-NMR. The WS fraction contains between 5.5 and 6.2 mmol/g of carbonyl groups, between 0.4 and 1.0 mmol/g of carboxylic acid groups, between 1.2 and 1.8 mmol/g phenolic -OH, and between 6.0 and 7.9 mmol/g of aliphatic alcohol groups. Translation into weight fractions of the WS was done by supposing surrogate structures for the water soluble phenols, carbonyl and carboxyl groups and we estimated the content of WS phenols (21-27 wt. %), carbonyl (5-14 wt.%), and carboxyl (0-4 wt.%). Together with the total carbohydrates (23-27 wt.%), this approach leads to > 90 wt. % of the WS material in the bio-oils being quantified. We speculate the larger portion of the difference between the total carbohydrates and hydrolysable sugars is the missing furanic fraction. Further refinement of the suggested methods and development of separation schemes to obtain and

Hydrothermal degradation of cellulosic matter to sugars and their fermentative conversion to protein

International Nuclear Information System (INIS)

Bobleter, O.; Niesner, R.; Roehr, M.

1976-01-01

For the hydrothermal degradation of cellulosic matter, an apparatus was developed in which water is used as extraction medium. Samples, 0.15 g each, of pure cellulose (filter paper), natural straw, and 14 C-labeled straw were treated at temperatures of between 200 and 275 0 C. Of the inserted cellulose, 65.7 percent was recovered at the optimum temperature as sugars and hydroxymethylfurfural. It was possible to degrade the straw selectively: at lower temperatures, the hemicellulose part of the plant matter was converted to xylose and arabinose; and then at higher temperatures, the cellulose was converted to glucose and cellobiose. At the same time, a certain amount of the sugars was transformed to furfural compounds. The growth behavior of the yeast Candida utilis (strain Weissenbach) was analyzed, using cellobiose, xylose, and glucose (standard) as carbon sources. The growth curves applying cellobiose were nearly identical to those of glucose. Xylose showed lower productivity than the hexoses. The main products of the hydrothermal degradation can, therefore, be used favorably as nutritive substances for this protein-producing yeast

Surface properties of heat-induced soluble soy protein aggregates of different molecular masses.

Science.gov (United States)

Guo, Fengxian; Xiong, Youling L; Qin, Fang; Jian, Huajun; Huang, Xiaolin; Chen, Jie

2015-02-01

Suspensions (2% and 5%, w/v) of soy protein isolate (SPI) were heated at 80, 90, or 100 °C for different time periods to produce soluble aggregates of different molecular sizes to investigate the relationship between particle size and surface properties (emulsions and foams). Soluble aggregates generated in these model systems were characterized by gel permeation chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Heat treatment increased surface hydrophobicity, induced SPI aggregation via hydrophobic interaction and disulfide bonds, and formed soluble aggregates of different sizes. Heating of 5% SPI always promoted large-size aggregate (LA; >1000 kDa) formation irrespective of temperature, whereas the aggregate size distribution in 2% SPI was temperature dependent: the LA fraction progressively rose with temperature (80→90→100 °C), corresponding to the attenuation of medium-size aggregates (MA; 670 to 1000 kDa) initially abundant at 80 °C. Heated SPI with abundant LA (>50%) promoted foam stability. LA also exhibited excellent emulsifying activity and stabilized emulsions by promoting the formation of small oil droplets covered with a thick interfacial protein layer. However, despite a similar influence on emulsion stability, MA enhanced foaming capacity but were less capable of stabilizing emulsions than LA. The functionality variation between heated SPI samples is clearly related to the distribution of aggregates that differ in molecular size and surface activity. The findings may encourage further research to develop functional SPI aggregates for various commercial applications. © 2015 Institute of Food Technologists®

Palatability of water-soluble extracts of protein sources and replacement of fishmeal by a selected mixture of protein sources for juvenile turbot ( Scophthalmus maximus)

Science.gov (United States)

Dong, Chun; He, Gen; Mai, Kangsen; Zhou, Huihui; Xu, Wei

2016-06-01

Poor palatability is a limiting factor for replacing fishmeal with other protein sources in aquaculture. The water-soluble molecules with low molecular weights are the major determinants of the palatability of diets. The present study was conducted to investigate the palatability of water-soluble extracts from single protein source (single extract pellets) and the mixture of these extracts with different proportions (blended extract pellets) in juvenile turbot ( Scophthalmus maximus). Then according to the palatability of blended extract pellets, an optimal mixture proportion was selected, and a new protein source made from raw protein materials with the selected proportion was formulated to replace fishmeal. Summarily, the palatability of single extract pellets for turbot was descendent from fishmeal to pet-food grade poultry by-product meal, wheat gluten meal, soybean meal, peanut meal, meat and bone meal, and corn gluten meal. Subsequently, according to the palatability of single extract pellets, 52 kinds of blended extract pellets were designed to test their palatability. The results showed that the pellets presented remarkably different palatability, and the optimal one was diet 52 (wheat gluten meal: pet-food grade poultry by-product meal: meat and bone meal: corn gluten meal = 1:6:1:2). The highest ingestion ratio (the number of pellets ingested/the number of pellets fed) was 0.73 ± 0.03, which was observed in Diet 52. Then five isonitrogenous (52% crude protein) and isocaloric (20 kJ g-1 gross energy) diets were formulated by replacing 0 (control), 35%, 50%, 65% and 80% of fishmeal with No.52 blending proportion. After a 10-weeks feeding trial, a consistent feed intake was found among all replacement treatments. Replacement level of fishmeal up to 35% did not significantly influence final body weight, specific growth rate, feed efficiency ratio, and protein efficiency ratio of turbot. Therefore, the water-soluble extracts of protein sources play an

Study the Effect of Different Phosphorus Fertilizers on Physiological Characteristic of Photosynthetic Pigments and Soluble Sugars of Safflower under Water Deficit Condition

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S Heshmati

2016-10-01

capacity. The sub-factors were the six treatments resulting from three levels of phosphate chemical fertilizer (0, 50, and 100 kg ha-1 Triple Super Phosphate, each at two levels of Barvar-2 bio-fertilizer (with and without inoculation with Barvar-2. The applied biological fertilizer was in the type of the bacteria which release the phosphorus from soil components and neutralize the soil pH. The commercial name of the biologic fertilizer is Barvar-2. The effective gradients of the biological phosphorus fertilizer is comprised of two bacteria strains of p5 (Bacillus lentus and p13 (Pseudomonas potida with 108 cfu (colony forming units which have been screened from soil bacteria populations. The bacteria strain p5 (Bacillus lentus dissolves P from soil mineral compounds while p13 (Pseudomonas potida separates P from soil organic compounds by exerting a variety of phosphatase enzymes. Results and Discussion The results showed that the highest amount of chlorophyll a, b and total chlorophyll obtained with application of chemical phosphorus by 100 kg ha-1 Triple Super Phosphate followed by inoculation with Barvar-2 under water deficit condition at vegetation and flowering stages. At drought stress in flowering stage, use of high level of phosphorus without inoculation with Barvar-2, had highest effect in terms of increasing amount of chlorophyll a/b ratio, but at treatment of inoculation with Barvar-2, the highest amount of chlorophyll a/ b ratio obtained with low level of chemical phosphorus by 50 kg ha-1 Triple Super Phosphate. The means comparison showed that highest amount of carotenoids, fv/fm, anthocyanin, flavonoids and soluble sugars obtained with application of chemical phosphorus by 50 kg ha-1 Triple Super Phosphate without Barvar-2 under water stress condition at vegetation and flowering stages. The inoculation with Barvar-2 followed by chemical phosphorus by 50 kg ha-1 Triple Super Phosphate significantly increased the amount of anthocyanin and soluble sugars of leaf

Leaf chemical composition of twenty-one Populus hybrid clones grown under intensive culture

Science.gov (United States)

Richard E. Dickson; Philip R. Larson

1976-01-01

Leaf material from 21 nursery-grown Populus hybrid clones was analyzed for three nitrogen fractions (total N, soluble protein, and soluble amino acids) and three carbhydrate fractions (reducing sugars, total soluble sugars, and total nonstructural carbohydrates-TNC). In addition, nursery-grown green ash and silver maple, field-grown bigtooth and trembling aspen, and...

Sialic Acid Binding Properties of Soluble Coronavirus Spike (S1 Proteins: Differences between Infectious Bronchitis Virus and Transmissible Gastroenteritis Virus

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Christine Winter

2013-07-01

Full Text Available The spike proteins of a number of coronaviruses are able to bind to sialic acids present on the cell surface. The importance of this sialic acid binding ability during infection is, however, quite different. We compared the spike protein of transmissible gastroenteritis virus (TGEV and the spike protein of infectious bronchitis virus (IBV. Whereas sialic acid is the only receptor determinant known so far for IBV, TGEV requires interaction with its receptor aminopeptidase N to initiate infection of cells. Binding tests with soluble spike proteins carrying an IgG Fc-tag revealed pronounced differences between these two viral proteins. Binding of the IBV spike protein to host cells was in all experiments sialic acid dependent, whereas the soluble TGEV spike showed binding to APN but had no detectable sialic acid binding activity. Our results underline the different ways in which binding to sialoglycoconjugates is mediated by coronavirus spike proteins.

Effect of enzymatic treatment of extracted sunflower proteins on solubility, amino acid composition, and surface activity.

Science.gov (United States)

Conde, José Miñones; Escobar, María del Mar Yust; Pedroche Jiménez, Justo J; Rodríguez, Francisco Millán; Rodríguez Patino, Juan M

2005-10-05

Industrial proteins from agriculture of either animal or vegetable origin, including their peptide derivatives, are of great importance, from the qualitative and quantitative point of view, in food formulations (emulsions and foams). A fundamental understanding of the physical, chemical, and functional properties of these proteins is essential if the performance of proteins in foods is to be improved and if underutilized proteins, such as plant proteins (and their hydrolysates and peptides derivatives), are to be increasingly used in traditional and new processed food products (safe, high-quality, health foods with good nutritional value). In this contribution we have determined the main physicochemical characteristics (solubility, composition, and analysis of amino acids) of a sunflower protein isolate (SPI) and its hydrolysates with low (5.62%), medium (23.5%), and high (46.3%) degrees of hydrolysis. The hydrolysates were obtained by enzymatic treatment with Alcalase 2.4 L for DH 5.62 and 23.5% and with Alcalase 2.4 L and Flavorzyme 1000 MG sequentially for DH 46.3%. The protein concentration dependence on surface pressure (surface pressure isotherm), a measure of the surface activity of the products (SPI and its hydrolysates), was obtained by tensiometry. We have observed that the degree of hydrolysis has an effect on solubility, composition, and content of the amino acids of the SPI and its hydrolysates. The superficial activity and the adsorption efficiency were also affected by the degree of hydrolysis.

Small acid soluble proteins for rapid spore identification.

Energy Technology Data Exchange (ETDEWEB)

Branda, Steven S.; Lane, Todd W.; VanderNoot, Victoria A.; Jokerst, Amanda S.

2006-12-01

This one year LDRD addressed the problem of rapid characterization of bacterial spores such as those from the genus Bacillus, the group that contains pathogenic spores such as B. anthracis. In this effort we addressed the feasibility of using a proteomics based approach to spore characterization using a subset of conserved spore proteins known as the small acid soluble proteins or SASPs. We proposed developing techniques that built on our previous expertise in microseparations to rapidly characterize or identify spores. An alternative SASP extraction method was developed that was amenable to both the subsequent fluorescent labeling required for laser-induced fluorescence detection and the low ionic strength requirements for isoelectric focusing. For the microseparations, both capillary isoelectric focusing and chip gel electrophoresis were employed. A variety of methods were evaluated to improve the molecular weight resolution for the SASPs, which are in a molecular weight range that is not well resolved by the current methods. Isoelectric focusing was optimized and employed to resolve the SASPs using UV absorbance detection. Proteomic signatures of native wild type Bacillus spores and clones genetically engineered to produce altered SASP patterns were assessed by slab gel electrophoresis, capillary isoelectric focusing with absorbance detection as well as microchip based gel electrophoresis employing sensitive laser-induced fluorescence detection.

Sugar uptake and starch biosynthesis by slices of developing maize endosperm

International Nuclear Information System (INIS)

Felker, F.C.; Liu, Kangchien; Shannon, J.C.

1990-01-01

14 C-Sugar uptake and incorporation into starch by slices of developing maize (Zea mays L.) endosperm were examined and compared with sugar uptake by maize endosperm-derived suspension cultures. Rates of sucrose, fructose, and D- and L-glucose uptake by slices were similar, whereas uptake rates for these sugars differed greatly in suspension cultures. Concentration dependence of sucrose, fructose, and D-glucose uptake was biphasic (consisting of linear plus saturable components) with suspension cultures but linear with slices. These and other differences suggest that endosperm slices are freely permeable to sugars. After diffusion into the slices, sugars were metabolized and incorporated into starch. Starch synthesis, but not sugar accumulation, was greatly reduced by 2.5 millimolar p-chloromercuribenzenesulfonic acid and 0.1 millimolar carbonyl cyanide m-chlorophenylhydrazone. Starch synthesis was dependent on kernel age and incubation temperature, but not on external pH (5 through 8). Competing sugars generally did not affect the distribution of 14 C among the soluble sugars extracted from endosperm slices incubated in 14 C-sugars. Competing hexoses reduced the incorporation of 14 C into starch, but competing sucrose did not, suggesting that sucrose is not a necessary intermediate in starch biosynthesis. The bidirectional permeability of endosperm slices to sugars makes the characterization of sugar transport into endosperm slices impossible, however the model system is useful for experiments dealing with starch biosynthesis which occurs in the metabolically active tissue

[Sugar Chain Construction of Functional Natural Products Using Plant Glucosyltransferases].

Science.gov (United States)

Mizukami, Hajime

2015-01-01

Plant secondary product glycosyltransferases belong to family 1 of the glycosyltransferase superfamily and mediate the transfer of a glycosyl residue from activated nucleotide sugars to lipophilic small molecules, thus affecting the solubility, stability and pharmacological activities of the sugar-accepting compounds. The biotechnological application of plant glycosyltransferases in glycoside synthesis has attracted attention because enzymatic glycosylation offers several advantages over chemical methods, including (1) avoiding the use of harsh conditions and toxic catalysts, (2) providing strict control of regio-and stereo-selectivity and (3) high efficiency. This review describes the in vivo and in vitro glycosylation of natural organic compounds using glycosyltransferases, focusing on our investigation of enzymatic synthesis of curcumin glycosides. Our current efforts toward functional characterization of some glycosyltransferases involved in the biosynthesis of iridoids and crocin, as well as in the sugar chain elongation of quercetin glucosides, are described. Finally, I describe the relationship of the structure of sugar chains and the intestinal absorption which was investigated using chemoenzymatically synthesized quercetin glycosides.

Synthesis of Salt Soluble Proteins in Barley. Pulse-Labeling Study of Grain Filling in Liquid-Cultured Detached Spikes

DEFF Research Database (Denmark)

Giese, Nanna Henriette; Hejgaard, Jørn

1984-01-01

The accumulation of salt-soluble proteins in the endosperm of developing barley (Hordeum vulgare L.) grains was examined. Detached spikes of barley were cultured at different levels of nitrogen nutrition and pulse-labeled with [14C] sucrose at specific times after anthesis. Proteins were extracted...... to increased nitrogen nutrition. Two major components, β-amylase and protein Z in particular, had a synthesis profile almost identical to that of the endosperm storage protein, hordein....

Proteomic Profiling of Sugar Beet (Beta vulgaris Leaves during Rhizomania Compatible Interactions

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Kimberly M. Webb

2014-04-01

Full Text Available Rhizomania, caused by Beet necrotic yellow vein virus (BNYVV, severely impacts sugar beet (Beta vulgaris production throughout the world, and is widely prevalent in most production regions. Initial efforts to characterize proteome changes focused primarily on identifying putative host factors that elicit resistant interactions with BNYVV, but as resistance breaking strains become more prevalent, effective disease control strategies will require the application of novel methods based on better understanding of disease susceptibility and symptom development. Herein, proteomic profiling was conducted on susceptible sugar beet, infected with two strains of BNYVV, to clarify the types of proteins prevalent during compatible virus-host plant interactions. Total protein was extracted from sugar beet leaf tissue infected with BNYVV, quantified, and analyzed by mass spectrometry. A total of 203 proteins were confidently identified, with a predominance of proteins associated with photosynthesis and energy, metabolism, and response to stimulus. Many proteins identified in this study are typically associated with systemic acquired resistance and general plant defense responses. These results expand on relatively limited proteomic data available for sugar beet and provide the ground work for additional studies focused on understanding the interaction of BNYVV with sugar beet.

Pretreatment of Dried Distiller Grains with Solubles by Soaking in Aqueous Ammonia and Subsequent Enzymatic/Dilute Acid Hydrolysis to Produce Fermentable Sugars.

Science.gov (United States)

Nghiem, Nhuan P; Montanti, Justin; Kim, Tae Hyun

2016-05-01

Dried distillers grains with solubles (DDGS), a co-product of corn ethanol production in the dry-grind process, was pretreated by soaking in aqueous ammonia (SAA) using a 15 % w/w NH4OH solution at a solid/liquid ratio of 1:10. The effect of pretreatment on subsequent enzymatic hydrolysis was studied at two temperatures (40 and 60 °C) and four reaction times (6, 12, 24, and 48 h). Highest glucose yield of 91 % theoretical was obtained for the DDGS pretreated at 60 °C and 24 h. The solubilized hemicellulose in the liquid fraction was further hydrolyzed with dilute H2SO4 to generate fermentable monomeric sugars. The conditions of acid hydrolysis included 1 and 4 wt% acid, 60 and 120 °C, and 0.5 and 1 h. Highest yields of xylose and arabinose were obtained at 4 wt% acid, 120 °C, and 1 h. The fermentability of the hydrolysate obtained by enzymatic hydrolysis of the SAA-pretreated DDGS was demonstrated in ethanol fermentation by Saccharomyces cerevisiae. The fermentability of the hydrolysate obtained by consecutive enzymatic and dilute acid hydrolysis was demonstrated using a succinic acid-producing microorganism, strain Escherichia coli AFP184. Under the fermentation conditions, complete utilization of glucose and arabinose was observed, whereas only 47 % of xylose was used. The succinic acid yield was 0.60 g/g total sugar consumed.

A tetrodotoxin-binding protein in the hemolymph of shore crab Hemigrapsus sanguineus: purification and properties.

Science.gov (United States)

Nagashima, Yuji; Yamamoto, Kazuhiko; Shimakura, Kuniyoshi; Shiomi, Kazuo

2002-06-01

The shore crab Hemigrapsus sanguineus hemolymph contains soluble proteins that bind tetrodotoxin (TTX) and are responsible for high resistance of the crab to TTX. The TTX-binding protein was purified from the hemolymph by ultrafiltration, lectin affinity chromatography and gel filtration HPLC. The purified protein gave only one band in native-polyacrylamide gel electrophoresis (PAGE), confirming its homogeneity. Its molecular weight was estimated to be about 400k by gel filtration HPLC, while it was estimated to be about 82k under non-reducing conditions and about 72 and 82k under reducing conditions by SDS-PAGE, indicating that the TTX-binding protein was composed of at least two distinct subunits. The TTX-binding protein was an acidic glycoprotein with pI 3.5, abundant in Asp and Glu but absent in Trp, and contained 6% reducing sugar and 12% amino sugar. The protein selectively bound to TTX, with a neutralizing ability of 6.7 mouse unit TTX/mg protein, but not to paralytic shellfish poisoning toxins. However, its neutralizing activity was almost lost by treatments with enzymes (protease XIV, thermolysin, trypsin, amyloglucosidase and alpha-amylase) and denaturing agents (1% SDS, 1% dithiothreitol, 8 M urea and 6 M guanidine hydrochloride), suggesting the involvement of both proteinaceous and sugar moieties in the binding to TTX and the importance of the steric conformation of the TTX-binding protein. Copright 2002 Elsevier Science Ltd.

Spectroscopic studies on the interaction of a water-soluble cationic porphyrin with proteins

Science.gov (United States)

Ma, Hong-Min; Chen, Xin; Zhang, Nuo; Han, Yan-Yan; Wu, Dan; Du, Bin; Wei, Qin

2009-04-01

The interaction of a water-soluble cationic porphyrin, meso-tetrakis (4- N, N, N-trimethylanilinium) porphyrin (TMAP), with two proteins, bovine serum albumin (BSA) and human serum albumin (HSA), was studied by UV-vis absorption spectroscopy, fluorescence spectroscopy, fluorescence anisotropy and synchronous fluorescence spectroscopy at neutral aqueous solutions. Free base TMAP bound to proteins as monomers and no aggregation was observed. The binding of TMAP quenched the fluorescence of the protein. On the contrary, the fluorescence of TMAP was enhanced and the fluorescence anisotropy increased due to the binding. The direct static binding mechanism could account for the quenching by TMAP and the binding constants were calculated. TMAP showed a higher quenching efficiency and binding constant of HSA than BSA. The binding of TMAP had no obvious effect on the molecular conformation of the protein. There was only one binding site for TMAP and it was located on the surface of the protein molecule. Electrostatic force played an important role in the binding due to the opposite charges on porphyrin and the proteins.

Application of a PEG precipitation method for solubility screening: A tool for developing high protein concentration formulations

OpenAIRE

Li, Li; Kantor, Angela; Warne, Nicholas

2013-01-01

Previous publications demonstrated that the extrapolated solubility by polyethylene glycol (PEG) precipitation method (Middaugh et al., J Biol Chem 1979; 254:367–370; Juckes, Biochim Biophys Acta 1971; 229:535–546; Foster et al., Biochim Biophys Acta 1973; 317:505; Mahadevan and Hall, AIChE J 1990; 36:1517–1528; Stevenson and Hageman, Pharm Res 1995; 12:1671–1676) has a strong correlation to experimentally measured solubility of proteins. Here, we explored the utility of extrapolated solubili...

Subcellular localization and logistics of integral membrane protein biogenesis in Escherichia coli.

Science.gov (United States)

Bogdanov, Mikhail; Aboulwafa, Mohammad; Saier, Milton H

2013-01-01

Transporters catalyze entry and exit of molecules into and out of cells and organelles, and protein-lipid interactions influence their activities. The bacterial phosphoenolpyruvate: sugar phosphotransferase system (PTS) catalyzes transport-coupled sugar phosphorylation as well as nonvectorial sugar phosphorylation in the cytoplasm. The vectorial process is much more sensitive to the lipid environment than the nonvectorial process. Moreover, cytoplasmic micellar forms of these enzyme-porters have been identified, and non-PTS permeases have similarly been shown to exist in 'soluble' forms. The latter porters exhibit lipid-dependent activities and can adopt altered topologies by simply changing the lipid composition. Finally, intracellular membranes and vesicles exist in Escherichia coli leading to the following unanswered questions: (1) what determines whether a PTS permease catalyzes vectorial or nonvectorial sugar phosphorylation? (2) How do phospholipids influence relative amounts of the plasma membrane, intracellular membrane, inner membrane-derived vesicles and cytoplasmic micelles? (3) What regulates the route(s) of permease insertion and transfer into and between the different subcellular sites? (4) Do these various membranous forms have distinct physiological functions? (5) What methods should be utilized to study the biogenesis and interconversion of these membranous structures? While research concerning these questions is still in its infancy, answers will greatly enhance our understanding of protein-lipid interactions and how they control the activities, conformations, cellular locations and biogenesis of integral membrane proteins. Copyright © 2013 S. Karger AG, Basel.

GC-EI-MS identification data of neutral sugars of polysaccharides extracted from Zizyphus lotus fruit

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Khaoula Mkadmini Hammi

2018-06-01

Full Text Available Gas chromatography coupled to mass spectrometer (GC–MS was used to identify and to quantify neutral sugars that constitute the water soluble polysaccharides from Zizyphus lotus fruit. The trimethylsilyl (TMS method was successfully used for derivatization of the monosaccharides units of extracted polysaccharides that were released by hydrolysis method. Sugars were identified based on their retention times compared with those of standards and the NIST MS Spectral Library. All sugars were quantified in TIC (Total Ion Current mode using calibration curves. Data is related to “Optimization extraction of polysaccharide from Tunisian Zizyphus lotus fruit by response surface methodology: Composition and antioxidant activity” (Mkadmini Hammi et al., 2016 [1]. Keywords: Trimethylsilyl, Derivatization, GC–MS, Neutral sugar

Epicotyl dormancy of tree peony as an oil plant broken by cyanamide

Science.gov (United States)

Xu, Jiajie; Gong, Mingfu; Liu, Fang; Wu, Sanlin; Liu, Xiaojie; Zhang, Ya; Xu, Gaoyu

2018-04-01

This test materials is `feng Dan', an oil peony, or tree peony as an oil plant, growing in Yangtze river basin. Impact of cyanamide on oil peony epicotyl dormancy was represented with germination rate of peony feeds, a-amylase activity, soluble sugar content, soluble protein content and peroxidase (POD) activity. Results showed that hypocotyls' dormancy of peony seeds was significant breaken by 0.3% cyanamide concentration. Alpha-amylase activity, soluble sugar content, soluble protein content and POD activity in 0.3% cyanamide concentration treatment was significantly higher than other treatments. There was no significant difference between the rest treatments.

The nutritive value of condensed wheat distillers solubles for cattle.

Science.gov (United States)

De Boever, J L; Blok, M C; Millet, S; Vanacker, J; De Campeneere, S

2016-12-01

The chemical composition and the energy and protein value of five batches of condensed distillers solubles (CDS) originating from wheat were determined. The net energy for lactation (NEL) was derived from digestion coefficients obtained with sheep. The true protein digested in the small intestine (DVE) and the rumen degradable protein balance (OEB) were based on the rumen degradation rate (kd D ), the rumen undegradable fraction (U) and intestinal digestibility of undegraded protein (%DVBE) predicted by regression equations derived from a data set of 28 protein feeds with kd D , U and %DVBE determined in situ. The CDS is a by-product with a high, but very variable CP content (238 to 495 g/kg DM). The CP contained on average 81% amino acids, with glutamine as main component (on average 21.8% of CP) and a relatively good lysine proportion (3.0%). Further, CDS contains quite a lot of crude fat (mean±SD: 71±14 g/kg DM), glycerol (95±52 g/kg DM) and sugars (123±24 g/kg DM) resulting in a high organic matter digestibility (88.6±3.0%) and high NEL content (8.3±0.4 MJ/kg DM). The protein value showed a large variation, with DVE ranging from 122 to 244 g/kg DM and OEB from 50 to 204 g/kg DM. Wheat CDS is a rich source of minerals and trace elements with exception of calcium.

Bacterial CpG-DNA activates dendritic cells in vivo: T helper cell-independent cytotoxic T cell responses to soluble proteins.

Science.gov (United States)

Sparwasser, T; Vabulas, R M; Villmow, B; Lipford, G B; Wagner, H

2000-12-01

Receptors for conserved molecular patterns associated with microbial pathogens induce synthesis of co-stimulatory molecules and cytokines in immature dendritic cells (DC), as do antigen-reactive CD4 T helper cells via CD40 signaling. Once activated, antigen-presenting DC may activate CD8 T cell responses in a T helper cell-independent fashion. Using immunostimulatory CpG-oligonucleotides (ODN) mimicking bacterial CpG-DNA, we tested whether CpG-DNA bypasses the need for T helper cells in CTL responses towards proteins by directly activating antigen-presenting DC to transit into professional APC. We describe that immature DC in situ constitutively process soluble proteins and generate CD8 T cell determinants yet CD8 T cell responses remain abortive. Induction of primary antigen-specific CD8 cytotoxic T lymphocyte (CTL)-mediated responses becomes initiated in wild-type as well as T helper cell-deficient mice, provided soluble protein and CpG-ODN are draining into the same lymph node. Specifically we show that CpG-ODN trigger antigen-presenting immature DC within the draining lymph node to acutely up-regulate co-stimulatory molecules and produce IL-12. These results provide new insights for generating in vivo efficient CTL responses to soluble proteins which may influence vaccination strategies.

Soluble carbohydrates in cereal (wheat, rye, triticale seed after storage under accelerated ageing conditions

Directory of Open Access Journals (Sweden)

Agnieszka I. Piotrowicz-Cieślak

2011-01-01

Full Text Available Germinability and the content of soluble carbohydrates were analysed in cereal seed (winter rye, cv. Warko; spring wheat, cv. Santa; hexaploid winter triticale, cv. Fidelio and cv. Woltario. Seed moisture content (mc was equilibrated over silica gel to 0.08 g H2O/g dry mass and stored in a desiccator at 20oC for up to 205 weeks or were equilibrated to mc 0.06, 0.08 or 0.10 g H2O/g dm and subjected to artificial aging at 35oC in air-tight laminated aluminium foil packages for 205 weeks. It was shown that the rate of seed aging depended on the species and seed moisture content. The fastest decrease of germinability upon storage was observed in seed with the highest moisture level. Complete germinability loss for winter rye, winter triticale cv. Fidelio, winter triticale cv. Woltario and spring wheat seed with mc 0.10 g H2O/g dm3 occurred after 81, 81, 101 and 133 weeks, respectively. Fructose, glucose, galactose, myo-inositol, sucrose, galactinol, raffinose, stachyose and verbascose were the main soluble carbohydrates found in the seed. The obtained data on the contents of specific sugars and the composition of soluble sugars fraction in seed of rye, wheat and triticale did not corroborate any profound effect of reducing sugars, sucrose and oligosaccharides on seed longevity.

Heat-induced alterations in cashew allergen solubility and IgE binding

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Christopher P. Mattison

Full Text Available Cashew nuts are an increasingly common cause of food allergy. We compare the soluble protein profile of cashew nuts following heating. SDS-PAGE indicate that heating can alter the solubility of cashew nut proteins. The 11S legumin, Ana o 2, dominates the soluble protein content in ready to eat and mildly heated cashew nuts. However, we found that in dark-roasted cashew nuts, the soluble protein profile shifts and the 2S albumin Ana o 3 composes up to 40% of the soluble protein. Analysis of trypsin-treated extracts by LC/MS/MS indicate changes in the relative number and intensity of peptides. The relative cumulative intensity of the 5 most commonly observed Ana o 1 and 2 peptides are altered by heating, while those of the 5 most commonly observed Ana o 3 peptides remaine relatively constant. ELISA experiments indicate that there is a decrease in rabbit IgG and human serum IgE binding to soluble cashew proteins following heating. Our findings indicate that heating can alter the solubility of cashew allergens, resulting in altered IgE binding. Our results support the use of both Ana o 2 and Ana o 3 as potential cashew allergen diagnostic targets. Keywords: Cashew nut, Food allergy, Immunoglobulin E, Mass-spectrometry, Peptide, Solubility

SILAGE CANE SUGAR ADDED WITH DRIED BREWER

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W. J. R. Castro

2015-02-01

Full Text Available The objective of this experiment was to evaluate the fermentative parameters and chemical composition of silage cane sugar added with residue dried brewery. The experimental design was completely randomized with four treatments and four replications: 100% cane sugar; 90% of cane sugar + 10% residue dried brewer; 80% of cane sugar + 20% residue dried brewer and 70% cane sugar + 30% dried brewer based on natural matter, composed silages. The sugar cane was chopped in a stationary machine with forage particle size of approximately 2 cm, and homogenized manually with the additives. For storage chopped fresh weight were used in experimental silos capacity of about 4 liters. The results showed that the contents of dry matter and crude protein showed positive linear (P0.05 with mean value of 3.81, while for ether extract and ash results were positive linear (P0.05 for N ammonia presented average value of 4.18. It is concluded that the addition of brewer dehydrated improves the fermentation process of silage cane sugar, in addition to improving their nutritional characteristics.

Plasma soluble prion protein, a potential biomarker for sport-related concussions: a pilot study.

Science.gov (United States)

Pham, Nam; Akonasu, Hungbo; Shishkin, Rhonda; Taghibiglou, Changiz

2015-01-01

Sport-related mild traumatic brain injury (mTBI) or concussion is a significant health concern to athletes with potential long-term consequences. The diagnosis of sport concussion and return to sport decision making is one of the greatest challenges facing health care clinicians working in sports. Blood biomarkers have recently demonstrated their potential in assisting the detection of brain injury particularly, in those cases with no obvious physical injury. We have recently discovered plasma soluble cellular prion protein (PrP(C)) as a potential reliable biomarker for blast induced TBI (bTBI) in a rodent animal model. In order to explore the application of this novel TBI biomarker to sport-related concussion, we conducted a pilot study at the University of Saskatchewan (U of S) by recruiting athlete and non-athlete 18 to 30 year-old students. Using a modified quantitative ELISA method, we first established normal values for the plasma soluble PrP(C) in male and female students. The measured plasma soluble PrP(C) in confirmed concussion cases demonstrated a significant elevation of this analyte in post-concussion samples. Data collected from our pilot study indicates that the plasma soluble PrP(C) is a potential biomarker for sport-related concussion, which may be further developed into a clinical diagnostic tool to assist clinicians in the assessment of sport concussion and return-to-play decision making.

The synthesis of a water-soluble derivative of rutin as an antiradical agent

Energy Technology Data Exchange (ETDEWEB)

Pedriali, Carla Aparecida; Fernandes, Adjaci Uchoa [Universidade de Sao Paulo (USP), SP (Brazil). Inst. de Quimica. Dept. de Bioquimica]. E-mail: capedriali@hotmail.com; Bernusso, Leandra de Cassia; Polakiewicz, Bronislaw [Universidade de Sao Paulo (USP), SP (Brazil). Faculdade de Ciencias Farmaceuticas. Dept. de Tecnologia Bioquimico-Farmaceutica

2008-07-01

The purpose of this study was to synthesize a water-soluble derivative of rutin (compound 2) by introducing carboxylate groups on rutin's sugar moiety. The rutin derivative showed an almost 100-fold solubility increase in water. The antiradical capacity of compound 2 was evaluated using the luminol/AAPH system, and the derivative's activity was 1.5 times greater than that of Trolox. Despite the derivative's high solubility in water (log P = -1.13), lipid peroxidation of brain homogenate membranes was very efficiently inhibited (inhibition values were only 19% lower than the inhibition values of rutin). (author)

The synthesis of a water-soluble derivative of rutin as an antiradical agent

International Nuclear Information System (INIS)

Pedriali, Carla Aparecida; Fernandes, Adjaci Uchoa; Bernusso, Leandra de Cassia; Polakiewicz, Bronislaw

2008-01-01

The purpose of this study was to synthesize a water-soluble derivative of rutin (compound 2) by introducing carboxylate groups on rutin's sugar moiety. The rutin derivative showed an almost 100-fold solubility increase in water. The antiradical capacity of compound 2 was evaluated using the luminol/AAPH system, and the derivative's activity was 1.5 times greater than that of Trolox. Despite the derivative's high solubility in water (log P = -1.13), lipid peroxidation of brain homogenate membranes was very efficiently inhibited (inhibition values were only 19% lower than the inhibition values of rutin). (author)

Effect of phospholipid, detergent and protein-protein interaction on stability and phosphoenzyme isomerization of soluble sarcoplasmic reticulum Ca-ATPase.

Science.gov (United States)

Vilsen, B; Andersen, J P

1987-12-30

The purpose of the present study was to elucidate the separate roles of lipid, detergent and protein-protein interaction for stability and catalytic properties of sarcoplasmic reticulum Ca-ATPase solubilized in the non-ionic detergent octa(ethylene glycol) monododecyl ether (C12E8). The use of large-zone high-performance liquid chromatography permitted us to define the self-association state of Ca-ATPase peptide at various detergent, phospholipid and protein concentrations, and also during enzymatic turnover with ATP. Conditions were established for monomerization of Ca-ATPase in the presence of a high concentration of phospholipid relative to detergent. The lipid-saturated monomeric preparation was relatively resistant to inactivation in the absence of Ca2+, whereas delipidated enzyme in monomeric or in oligomeric form was prone to inactivation. Kinetics of phosphoenzyme turnover were examined in the presence and absence of Mg2+. Dephosphorylation rates were sensitive to Mg2+, irrespective of whether the peptide was present in soluble monomeric form or was membrane-bound. C12E8-solubilized monomer without added phospholipid was, however, characterized by a fast initial phase of dephosphorylation in the absence of Mg2+. This was not observed with monomer saturated with phospholipid or with monomer solubilized in myristoylglycerophosphocholine or deoxycholate. The mechanism underlying this difference was shown to be a C12E8-induced acceleration of conversion of ADP-sensitive phosphoenzyme (E1P) to ADP-insensitive phosphoenzyme (E2P). The phosphoenzyme isomerization rate was also found to be enhanced by low-affinity binding of ATP. This was demonstrated both in membrane-bound and in soluble monomeric Ca-ATPase. Our results indicate that a single peptide chain constitutes the target for modulation of phosphoenzyme turnover by Mg2+ and ATP, and that detergent effects, distinct from those arising from disruption of protein-protein contacts, are the major determinants of

Nonenzymatic glycosylation of bovine myelin basic protein

International Nuclear Information System (INIS)

Hitz, J.B.

1987-01-01

In the CNS myelin sheath the nonenzymatic glycosylation reaction (at the early stage of the Amadori product) occurs only with the myelin basic protein and not with the other myelin proteins. This was observed in isolated bovine myelin by in vitro incubation with [ 14 C]-galactose and [ 14 C]-glucose. The respective in-vitro incorporation rates for purified bovine myelin basic protein with D-galactose, D-glucose and D-mannose were 7.2, 2.4 and 2.4 mmoles/mole myelin basic protein per day at 37 0 C. A more rapid, HPLC method was devised and characterized to specifically analyze for the Amadori product. The HPLC method was correlated to the [ 14 C]-sugar incorporation method for myelin basic protein under a set of standard reaction conditions using [ 14 C]-glucose and [ 14 C]-mannose with HPLC values at 1/6 and 1/5 of the [ 14 C]-sugar incorporation method. A novel myelin basic protein purification step has been developed that yields a relativity proteolytic free preparation that is easy to work with, being totally soluble at a neutral pH. Nine new spots appear for a trypsinized glycosylated MBP in the paper peptide map of which eight correspond to positions of the [ 3 H]-labeled Amadori product in affinity isolated peptides. These studies provide a general characterization of and a structural basis for investigations on nonenzymatically glycosylated MBP as well as identifying MBP as the only nonenzymatically glycosylated protein in the CNS myelin sheath which may accumulate during aging, diabetes, and demyelinating diseases in general

Spray Drying of High Sugar Content Foods: Improving of Product Yield and Powder Properties

OpenAIRE

Mehmet Koç; Figen Kaymak-Ertekin

2016-01-01

Spray drying is the most preferred drying method to produce powdered food in the food industry and it is also widely used to convert sugar-rich liquid foods to a powder form. During and/or after spray drying process of sugar-rich products, undesirable situation was appeared such as stickiness, high moisture affinity (hygroscopicity) and low solubility due to low molecular weight monosaccharides that found naturally in the structure. The basis of these problems was formed by low glass transiti...

Current knowledge on agarolytic enzymes and the industrial potential of agar-derived sugars.

Science.gov (United States)

Yun, Eun Ju; Yu, Sora; Kim, Kyoung Heon

2017-07-01

Agar is a major cell wall carbohydrate of red macroalgae (Rhodophyta). Sugars derived from agar, such as agarooligosaccharides (AOSs), neoagarooligosaccharides (NAOSs), neoagarobiose (NAB), and 3,6-anhydro-L-galactose (L-AHG), possess various physiological activities. These agar-derived sugars can be produced by hydrolysis using chemicals or agarolytic enzymes. Despite the industrial potential of agar-derived sugars, their application has been hampered mainly due to the absence of efficient processes for the liquefaction and saccharification of agar. In this review, we have focused on strategies for producing high value-added sugars from agarose via chemical or enzymatic liquefaction and enzymatic saccharification. The liquefaction of agarose is a key step for preventing gelling and increasing the solubility of agarose in water by prehydrolyzing agarose into AOSs or NAOSs. For the industrial use of agar-derived sugars, AOS, NAOS, NAB, and L-AHG can be used as functional biomaterials owing to their physiological activities such as antiinflammation, skin whitening, and moisturizing. Recently, it was reported that AHG could be considered as a new anticariogenic sugar to replace xylitol. This review provides a comprehensive overview of processes for the hydrolysis of agar or agarose to produce high value-added sugars and the industrial application of these sugars.

Lipid-protein nanodiscs for cell-free production of integral membrane proteins in a soluble and folded state: comparison with detergent micelles, bicelles and liposomes.

Science.gov (United States)

Lyukmanova, E N; Shenkarev, Z O; Khabibullina, N F; Kopeina, G S; Shulepko, M A; Paramonov, A S; Mineev, K S; Tikhonov, R V; Shingarova, L N; Petrovskaya, L E; Dolgikh, D A; Arseniev, A S; Kirpichnikov, M P

2012-03-01

Production of integral membrane proteins (IMPs) in a folded state is a key prerequisite for their functional and structural studies. In cell-free (CF) expression systems membrane mimicking components could be added to the reaction mixture that promotes IMP production in a soluble form. Here lipid-protein nanodiscs (LPNs) of different lipid compositions (DMPC, DMPG, POPC, POPC/DOPG) have been compared with classical membrane mimicking media such as detergent micelles, lipid/detergent bicelles and liposomes by their ability to support CF synthesis of IMPs in a folded and soluble state. Three model membrane proteins of different topology were used: homodimeric transmembrane (TM) domain of human receptor tyrosine kinase ErbB3 (TM-ErbB3, 1TM); voltage-sensing domain of K(+) channel KvAP (VSD, 4TM); and bacteriorhodopsin from Exiguobacterium sibiricum (ESR, 7TM). Structural and/or functional properties of the synthesized proteins were analyzed. LPNs significantly enhanced synthesis of the IMPs in a soluble form regardless of the lipid composition. A partial disintegration of LPNs composed of unsaturated lipids was observed upon co-translational IMP incorporation. Contrary to detergents the nanodiscs resulted in the synthesis of ~80% active ESR and promoted correct folding of the TM-ErbB3. None of the tested membrane mimetics supported CF synthesis of correctly folded VSD, and the protocol of the domain refolding was developed. The use of LPNs appears to be the most promising approach to CF production of IMPs in a folded state. NMR analysis of (15)N-Ile-TM-ErbB3 co-translationally incorporated into LPNs shows the great prospects of this membrane mimetics for structural studies of IMPs produced by CF systems. Copyright © 2011 Elsevier B.V. All rights reserved.

Enhanced Soluble Protein and Biochemical Methane Potential of Apple Biowaste by Different Pretreatment

Science.gov (United States)

Tulun, Şevket; Bilgin, Melayib

2018-05-01

The purpose of this research is to evaluate the anaerobic digestion of apple pomace waste in terms of pretreatment. In this study, the main pretreatment strategies for apple pomace include: ultrasound (35 and 53 kHz), thermal and chemical (pH 5 and 10). For each pretreatment method four different temperatures are selected as 25, 40, 50, and 60 °C, and operation times are selected as 5th, 15th, 30th, and 45th minutes. The effects on pretreatment were investigated by measuring changes in the soluble protein concentrations of pretreated wastes and the enhanced anaerobic digestion was investigated by using the biochemical methane potential (BMP) assay. The soluble proteins of ultrasonic (35 kHz at 60 °C, 45th min), ultrasonic (53 kHz at 60 °C, 45th min), chemical (pH 5 at 60 °C, 5th min), chemical (pH 10 at 60 °C, 30th min) and thermal chemical (40 °C, 15th min) pretreatment apple pomace were 74.3, 75.6, 48.7, 85.5 and 58.6% higher, respectively. The results indicated that apple pomace treated with 53 kHz at 60 °C, 45th min had the highest biogas yield of 1519 mL CH4/g VSS.day after anaerobic digestion, which was on average 40.9% higher than raw pomace.

Enhanced Soluble Protein and Biochemical Methane Potential of Apple Biowaste by Different Pretreatment

Science.gov (United States)

Tulun, Şevket; Bilgin, Melayib

2018-01-01

The purpose of this research is to evaluate the anaerobic digestion of apple pomace waste in terms of pretreatment. In this study, the main pretreatment strategies for apple pomace include: ultrasound (35 and 53 kHz), thermal and chemical (pH 5 and 10). For each pretreatment method four different temperatures are selected as 25, 40, 50, and 60 °C, and operation times are selected as 5th, 15th, 30th, and 45th minutes. The effects on pretreatment were investigated by measuring changes in the soluble protein concentrations of pretreated wastes and the enhanced anaerobic digestion was investigated by using the biochemical methane potential (BMP) assay. The soluble proteins of ultrasonic (35 kHz at 60 °C, 45th min), ultrasonic (53 kHz at 60 °C, 45th min), chemical (pH 5 at 60 °C, 5th min), chemical (pH 10 at 60 °C, 30th min) and thermal chemical (40 °C, 15th min) pretreatment apple pomace were 74.3, 75.6, 48.7, 85.5 and 58.6% higher, respectively. The results indicated that apple pomace treated with 53 kHz at 60 °C, 45th min had the highest biogas yield of 1519 mL CH4/g VSS.day after anaerobic digestion, which was on average 40.9% higher than raw pomace.

Wheat germ cell-free expression: Two detergents with a low critical micelle concentration allow for production of soluble HCV membrane proteins.

Science.gov (United States)

Fogeron, Marie-Laure; Badillo, Aurélie; Jirasko, Vlastimil; Gouttenoire, Jérôme; Paul, David; Lancien, Loick; Moradpour, Darius; Bartenschlager, Ralf; Meier, Beat H; Penin, François; Böckmann, Anja

2015-01-01

Membrane proteins are notoriously difficult to express in a soluble form. Here, we use wheat germ cell-free expression in the presence of various detergents to produce the non-structural membrane proteins 2, 4B and 5A of the hepatitis C virus (HCV). We show that lauryl maltose neopentyl glycol (MNG-3) and dodecyl octaethylene glycol ether (C12E8) detergents can yield essentially soluble membrane proteins at detergent concentrations that do not inhibit the cell-free reaction. This finding can be explained by the low critical micelle concentration (CMC) of these detergents, which keeps the monomer concentrations low while at the same time providing the necessary excess of detergent concentration above CMC required for full target protein solubilization. We estimate that a tenfold excess of detergent micelles with respect to the protein concentration is sufficient for solubilization, a number that we propose as a guideline for detergent screening assays. Copyright © 2014 Elsevier Inc. All rights reserved.

Design of Fusion Proteins for Efficient and Soluble Production of Immunogenic Ebola Virus Glycoprotein in Escherichia coli.

Science.gov (United States)

Ji, Yang; Lu, Yuan; Yan, Yishu; Liu, Xinxin; Su, Nan; Zhang, Chong; Bi, Shengli; Xing, Xin-Hui

2018-03-03

The Ebola hemorrhagic fever caused by Ebola virus is an extremely dangerous disease, and effective therapeutic agents are still lacking. Platforms for the efficient production of vaccines are crucial to ensure quick response against an Ebola virus outbreak. Ebola virus glycoprotein (EbolaGP) on the virion surface is responsible for membrane binding and virus entry, thus becoming the key target for vaccine development. However, heterologous expression of this protein still faces engineering challenges such as low production levels and insoluble aggregation. Here, the authors design and compare various fusion strategies, attaching great importance to the solubility-enhancing effect, and tag removal process. It is found that a C-terminal intein-based tag greatly enhances the solubility of EbolaGP and allows one-step chromatographic purification of the untagged EbolaGP through thiol-catalyzed self-cleavage. The purified untagged EbolaGP alone or with Freund's adjuvant are highly immunogenic, as confirmed in a mouse model. Consequently, the present study puts forward a new strategy for the efficient and soluble expression of untagged immunogenic EbolaGP. The intein-based protein fusion approach may be of importance for the large-scale production of Ebola virus subunit vaccine. © 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A physicochemical study of sugar palm (Arenga Pinnata) starch films plasticized by glycerol and sorbitol

Science.gov (United States)

Poeloengasih, Crescentiana D.; Pranoto, Yudi; Hayati, Septi Nur; Hernawan, Rosyida, Vita T.; Prasetyo, Dwi J.; Jatmiko, Tri H.; Apriyana, Wuri; Suwanto, Andri

2016-02-01

The present work explores the physicochemical characteristics of sugar palm starch film for a potential hard capsule purpose. Sugar palm (Arenga pinnata) starch films were plasticized with glycerol or sorbitol in various concentrations (30% up to 50% w/w starch). Their effects on physicochemical properties of the films were investigated. The results showed that sugar palm starch was successfully developed as the main material of film using casting method. Incorporation of both glycerol or sorbitol affected the properties of films in different ways. It was found that thickness and solubility increased as plasticizer concentration increased, whereas retraction ratio, swelling degree and swelling thickness decreased with the increased plasticizer concentration.

A Versatile System for High-Throughput In Situ X-ray Screening and Data Collection of Soluble and Membrane-Protein Crystals

Energy Technology Data Exchange (ETDEWEB)

Broecker, Jana; Klingel, Viviane; Ou, Wei-Lin; Balo, Aidin R.; Kissick, David J.; Ogata, Craig M.; Kuo, Anling; Ernst, Oliver P.

2016-10-12

In recent years, in situ data collection has been a major focus of progress in protein crystallography. Here, we introduce the Mylar in situ method using Mylar-based sandwich plates that are inexpensive, easy to make and handle, and show significantly less background scattering than other setups. A variety of cognate holders for patches of Mylar in situ sandwich films corresponding to one or more wells makes the method robust and versatile, allows for storage and shipping of entire wells, and enables automated crystal imaging, screening, and goniometerbased X-ray diffraction data-collection at room temperature and under cryogenic conditions for soluble and membrane-protein crystals grown in or transferred to these plates. We validated the Mylar in situ method using crystals of the water-soluble proteins hen egg-white lysozyme and sperm whale myoglobin as well as the 7-transmembrane protein bacteriorhodopsin from Haloquadratum walsbyi. In conjunction with current developments at synchrotrons, this approach promises high-resolution structural studies of membrane proteins to become faster and more routine.

Proteomic analysis of Herbaspirillum seropedicae cultivated in the presence of sugar cane extract.

Science.gov (United States)

Cordeiro, Fabio Aparecido; Tadra-Sfeir, Michelle Zibetti; Huergo, Luciano Fernandes; de Oliveira Pedrosa, Fábio; Monteiro, Rose Adele; de Souza, Emanuel Maltempi

2013-03-01

Bacterial endophytes of the genus Herbaspirillum colonize sugar cane and can promote plant growth. The molecular mechanisms that mediate plant- H. seropedicae interaction are poorly understood. In this work, we used 2D-PAGE electrophoresis to identify H. seropedicae proteins differentially expressed at the log growth phase in the presence of sugar cane extract. The differentially expressed proteins were validated by RT qPCR. A total of 16 differential spots (1 exclusively expressed, 7 absent, 5 up- and 3 down-regulated) in the presence of 5% sugar cane extract were identified; thus the host extract is able to induce and repress specific genes of H. seropedicae. The differentially expressed proteins suggest that exposure to sugar cane extract induced metabolic changes and adaptations in H. seropedicae presumably in preparation to establish interaction with the plant.

Enhancing production and cytotoxic activity of polymeric soluble FasL-based chimeric proteins by concomitant expression of soluble FasL.

Directory of Open Access Journals (Sweden)

Aurore Morello

Full Text Available Membrane FasL is the natural trigger of Fas-mediated apoptosis. A soluble homotrimeric counterpart (sFasL also exists which is very weakly active, and needs oligomerization beyond its trimeric state to induce apoptosis. We recently generated a soluble FasL chimera by fusing the immunoglobulin-like domain of the leukemia inhibitory factor receptor gp190 to the extracellular region of human FasL, which enabled spontaneous dodecameric homotypic polymerization of FasL. This polymeric soluble human FasL (pFasL displayed anti-tumoral activity in vitro and in vivo without systemic cytotoxicity in mouse. In the present work, we focused on the improvement of pFasL, with two complementary objectives. First, we developed more complex pFasL-based chimeras that contained a cell-targeting module. Secondly, we attempted to improve the production and/or the specific activity of pFasL and of the cell-targeting chimeras. We designed two chimeras by fusing to pFasL the extracellular portions of the HLA-A2 molecule or of a human gamma-delta TCR, and analyzed the consequences of co-expressing these molecules or pFasL together with sFasL on their heterotopic cell production. This strategy significantly enhanced the production of pFasL and of the two chimeras, as well as the cytotoxic activity of the two chimeras but not of pFasL. These results provide the proof of concept for an optimization of FasL-based chimeric proteins for a therapeutic use.

Plasma soluble prion protein, a potential biomarker for sport-related concussions: a pilot study.

Directory of Open Access Journals (Sweden)

Nam Pham

Full Text Available Sport-related mild traumatic brain injury (mTBI or concussion is a significant health concern to athletes with potential long-term consequences. The diagnosis of sport concussion and return to sport decision making is one of the greatest challenges facing health care clinicians working in sports. Blood biomarkers have recently demonstrated their potential in assisting the detection of brain injury particularly, in those cases with no obvious physical injury. We have recently discovered plasma soluble cellular prion protein (PrP(C as a potential reliable biomarker for blast induced TBI (bTBI in a rodent animal model. In order to explore the application of this novel TBI biomarker to sport-related concussion, we conducted a pilot study at the University of Saskatchewan (U of S by recruiting athlete and non-athlete 18 to 30 year-old students. Using a modified quantitative ELISA method, we first established normal values for the plasma soluble PrP(C in male and female students. The measured plasma soluble PrP(C in confirmed concussion cases demonstrated a significant elevation of this analyte in post-concussion samples. Data collected from our pilot study indicates that the plasma soluble PrP(C is a potential biomarker for sport-related concussion, which may be further developed into a clinical diagnostic tool to assist clinicians in the assessment of sport concussion and return-to-play decision making.

Toxoplasma gondii: Biochemical and biophysical characterization of recombinant soluble dense granule proteins GRA2 and GRA6

International Nuclear Information System (INIS)

Bittame, Amina; Effantin, Grégory; Pètre, Graciane; Ruffiot, Pauline; Travier, Laetitia; Schoehn, Guy; Weissenhorn, Winfried; Cesbron-Delauw, Marie-France; Gagnon, Jean; Mercier, Corinne

2015-01-01

The most prominent structural feature of the parasitophorous vacuole (PV) in which the intracellular parasite Toxoplasma gondii proliferates is a membranous nanotubular network (MNN), which interconnects the parasites and the PV membrane. The MNN function remains unclear. The GRA2 and GRA6 proteins secreted from the parasite dense granules into the PV have been implicated in the MNN biogenesis. Amphipathic alpha-helices (AAHs) predicted in GRA2 and an alpha-helical hydrophobic domain predicted in GRA6 have been proposed to be responsible for their membrane association, thereby potentially molding the MMN in its structure. Here we report an analysis of the recombinant proteins (expressed in detergent-free conditions) by circular dichroism, which showed that full length GRA2 displays an alpha-helical secondary structure while recombinant GRA6 and GRA2 truncated of its AAHs are mainly random coiled. Dynamic light scattering and transmission electron microscopy showed that recombinant GRA6 and truncated GRA2 constitute a homogenous population of small particles (6–8 nm in diameter) while recombinant GRA2 corresponds to 2 populations of particles (∼8–15 nm and up to 40 nm in diameter, respectively). The unusual properties of GRA2 due to its AAHs are discussed. - Highlights: • Toxoplasma gondii: soluble GRA2 forms 2 populations of particles. • T. gondii: the dense granule protein GRA2 folds intrinsically as an alpha-helix. • T. gondii: monomeric soluble GRA6 forms particles of 6–8 nm in diameter. • T. gondii: monomeric soluble GRA6 is random coiled. • Unusual biophysical properties of the dense granule protein GRA2 from T. gondii

Toxoplasma gondii: Biochemical and biophysical characterization of recombinant soluble dense granule proteins GRA2 and GRA6

Energy Technology Data Exchange (ETDEWEB)

Bittame, Amina [CNRS, UMR 5163, 38042 Grenoble (France); Université Grenoble Alpes, 38042 Grenoble (France); Effantin, Grégory [Université Grenoble Alpes, Institut de Biologie Structurale (IBS), 38044 Grenoble (France); CNRS, IBS, 38044 Grenoble (France); CEA, IBS, 38044 Grenoble (France); Unit for Virus Host-Cell Interactions (UVHCI), UMI 3265 (UJF-EMBL-CNRS), 38027 Grenoble (France); Pètre, Graciane; Ruffiot, Pauline; Travier, Laetitia [CNRS, UMR 5163, 38042 Grenoble (France); Université Grenoble Alpes, 38042 Grenoble (France); Schoehn, Guy; Weissenhorn, Winfried [Université Grenoble Alpes, Institut de Biologie Structurale (IBS), 38044 Grenoble (France); CNRS, IBS, 38044 Grenoble (France); CEA, IBS, 38044 Grenoble (France); Unit for Virus Host-Cell Interactions (UVHCI), UMI 3265 (UJF-EMBL-CNRS), 38027 Grenoble (France); Cesbron-Delauw, Marie-France; Gagnon, Jean [CNRS, UMR 5163, 38042 Grenoble (France); Université Grenoble Alpes, 38042 Grenoble (France); Mercier, Corinne, E-mail: corinne.mercier@ujf-grenoble.fr [CNRS, UMR 5163, 38042 Grenoble (France); Université Grenoble Alpes, 38042 Grenoble (France)

2015-03-27

The most prominent structural feature of the parasitophorous vacuole (PV) in which the intracellular parasite Toxoplasma gondii proliferates is a membranous nanotubular network (MNN), which interconnects the parasites and the PV membrane. The MNN function remains unclear. The GRA2 and GRA6 proteins secreted from the parasite dense granules into the PV have been implicated in the MNN biogenesis. Amphipathic alpha-helices (AAHs) predicted in GRA2 and an alpha-helical hydrophobic domain predicted in GRA6 have been proposed to be responsible for their membrane association, thereby potentially molding the MMN in its structure. Here we report an analysis of the recombinant proteins (expressed in detergent-free conditions) by circular dichroism, which showed that full length GRA2 displays an alpha-helical secondary structure while recombinant GRA6 and GRA2 truncated of its AAHs are mainly random coiled. Dynamic light scattering and transmission electron microscopy showed that recombinant GRA6 and truncated GRA2 constitute a homogenous population of small particles (6–8 nm in diameter) while recombinant GRA2 corresponds to 2 populations of particles (∼8–15 nm and up to 40 nm in diameter, respectively). The unusual properties of GRA2 due to its AAHs are discussed. - Highlights: • Toxoplasma gondii: soluble GRA2 forms 2 populations of particles. • T. gondii: the dense granule protein GRA2 folds intrinsically as an alpha-helix. • T. gondii: monomeric soluble GRA6 forms particles of 6–8 nm in diameter. • T. gondii: monomeric soluble GRA6 is random coiled. • Unusual biophysical properties of the dense granule protein GRA2 from T. gondii.

Evaluation of a competitive enzyme-linked immunosorbent assay for measurements of soluble HLA-G protein.

Science.gov (United States)

Rasmussen, M; Dahl, M; Buus, S; Djurisic, S; Ohlsson, J; Hviid, T V F

2014-08-01

The human leukocyte antigen (HLA) class Ib molecule, HLA-G, has gained increased attention because of its assumed important role in immune regulation. The HLA-G protein exists in several soluble isoforms. Most important are the actively secreted HLA-G5 full-length isoform generated by alternative splicing retaining intron 4 with a premature stop codon, and the cleavage of full-length membrane-bound HLA-G1 from the cell surface, so-called soluble HLA-G1 (sHLA-G1). A specific and sensitive immunoassay for measurements of soluble HLA-G is mandatory for conceivable routine testing and research projects. We report a novel method, a competitive immunoassay, for measuring HLA-G5/sHLA-G1 in biological fluids. The sHLA-G immunoassay is based upon a competitive enzyme-linked immunosorbent assay (ELISA) principle. It includes a recombinant sHLA-G1 protein in complex with β2-microglobulin and a peptide as a standard, biotinylated recombinant sHLA-G1 as an indicator, and the MEM-G/9 anti-HLA-G monoclonal antibody (mAb) as the capture antibody. The specificity and sensitivity of the assay were evaluated. Testing with different recombinant HLA class I proteins and different anti-HLA class I mAbs showed that the sHLA-G immunoassay was highly specific. Optimal combinations of competitor sHLA-G1 and capture mAb concentrations were determined. Two versions of the assay were tested. One with a relatively wide dynamic range from 3.1 to 100.0 ng/ml, and another more sensitive version ranging from 1.6 to 12.5 ng/ml. An intra-assay coefficient of variation (CV) of 15.5% at 88 ng/ml and an inter-assay CV of 23.1% at 39 ng/ml were determined. An assay based on the competitive sHLA-G ELISA may be important for measurements of sHLA-G proteins in several conditions: assisted reproduction, organ transplantation, cancer, and certain pregnancy complications, both in research studies and possibly in the future also for clinical routine use. © 2014 John Wiley & Sons A/S. Published by John Wiley

The influence of the extraction parameters on the quality of dried sugar beet pulp

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Bogdanović Branislav V.

2013-01-01

Full Text Available Growing demand for better quality and cheaper products put on the need to industry to balance between the two kinds of needs, which are the reduction of production costs and to ensure a high quality product. This is the case with dried sugar beet pulp as a byproduct of sugar refining process. One of the possibilities to reduce energy consumption is savings in the process of thermal dehydration in the process of the conventional drying of pulp. Pulp drying is one third of total energy consumption in the sugar factory. Acidification of water extraction result in pulp that could be pressed better and in other hand in the juice with less non-sucrose compounds. Keeping the extraction process under certain conditions, directly affects on the quality of the extracted pulp and on effects of their further processing. This paper examines the impact of the extraction parameters of sugar from sugar beet and agents of acidification for water extraction (sulphurous acid, hydrogen peroxide on the quality of dried sugar beet pulp. Extraction of sugar from sugar beet cossettes by hydrogen peroxide at pH 5.5 to 8.5 does not cause changes in mechanical properties of sugar beet cossettes and in water retention coefficient, but increases the brightness of dried sugar beet pulp for about 5-7 units, which solubility in water does not depend on the applied concentration of extracting agents, but increases in dependence on the duration of the extraction. Sugar beet pulp extracted at pH 11.0 has reduced strength, they are highly hydrated, and under the force they lose their shape, binding a larger amount of water thus reducing the effect of their pressing. The changes in microstructure were probably accompanied by segmenting the linear parts of macromolecules and are reflected in the increase of water retention capacity and increase the solubility of dried sugar beet pulp for two to three times. Under the same conditions of extraction, sulphurous acid produces similar

Delayed Toxicity Associated with Soluble Anthrax Toxin Receptor Decoy-Ig Fusion Protein Treatment

Science.gov (United States)

Cote, Christopher; Welkos, Susan; Manchester, Marianne; Young, John A. T.

2012-01-01

Soluble receptor decoy inhibitors, including receptor-immunogloubulin (Ig) fusion proteins, have shown promise as candidate anthrax toxin therapeutics. These agents act by binding to the receptor-interaction site on the protective antigen (PA) toxin subunit, thereby blocking toxin binding to cell surface receptors. Here we have made the surprising observation that co-administration of receptor decoy-Ig fusion proteins significantly delayed, but did not protect, rats challenged with anthrax lethal toxin. The delayed toxicity was associated with the in vivo assembly of a long-lived complex comprised of anthrax lethal toxin and the receptor decoy-Ig inhibitor. Intoxication in this system presumably results from the slow dissociation of the toxin complex from the inhibitor following their prolonged circulation. We conclude that while receptor decoy-Ig proteins represent promising candidates for the early treatment of B. anthracis infection, they may not be suitable for therapeutic use at later stages when fatal levels of toxin have already accumulated in the bloodstream. PMID:22511955

Protein- protein interaction detection system using fluorescent protein microdomains

Science.gov (United States)

Waldo, Geoffrey S.; Cabantous, Stephanie

2010-02-23

The invention provides a protein labeling and interaction detection system based on engineered fragments of fluorescent and chromophoric proteins that require fused interacting polypeptides to drive the association of the fragments, and further are soluble and stable, and do not change the solubility of polypeptides to which they are fused. In one embodiment, a test protein X is fused to a sixteen amino acid fragment of GFP (.beta.-strand 10, amino acids 198-214), engineered to not perturb fusion protein solubility. A second test protein Y is fused to a sixteen amino acid fragment of GFP (.beta.-strand 11, amino acids 215-230), engineered to not perturb fusion protein solubility. When X and Y interact, they bring the GFP strands into proximity, and are detected by complementation with a third GFP fragment consisting of GFP amino acids 1-198 (strands 1-9). When GFP strands 10 and 11 are held together by interaction of protein X and Y, they spontaneous association with GFP strands 1-9, resulting in structural complementation, folding, and concomitant GFP fluorescence.

Identification of Proteins with Potential Osteogenic Activity Present in the Water-Soluble Matrix Proteins from Crassostrea gigas Nacre Using a Proteomic Approach

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Daniel V. Oliveira

2012-01-01

Full Text Available Nacre, when implanted in vivo in bones of dogs, sheep, mice, and humans, induces a biological response that includes integration and osteogenic activity on the host tissue that seems to be activated by a set of proteins present in the nacre water-soluble matrix (WSM. We describe here an experimental approach that can accurately identify the proteins present in the WSM of shell mollusk nacre. Four proteins (three gigasin-2 isoforms and a cystatin A2 were for the first time identified in WSM of Crassostrea gigas nacre using 2DE and LC-MS/MS for protein identification. These proteins are thought to be involved in bone remodeling processes and could be responsible for the biocompatibility shown between bone and nacre grafts. These results represent a contribution to the study of shell biomineralization process and opens new perspectives for the development of new nacre biomaterials for orthopedic applications.

Small surfactant-like peptides can drive soluble proteins into active aggregates

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Zhou Bihong

2012-01-01

Full Text Available Abstract Background Inactive protein inclusion bodies occur commonly in Escherichia coli (E. coli cells expressing heterologous proteins. Previously several independent groups have found that active protein aggregates or pseudo inclusion bodies can be induced by a fusion partner such as a cellulose binding domain from Clostridium cellulovorans (CBDclos when expressed in E. coli. More recently we further showed that a short amphipathic helical octadecapeptide 18A (EWLKAFYEKVLEKLKELF and a short beta structure peptide ELK16 (LELELKLKLELELKLK have a similar property. Results In this work, we explored a third type of peptides, surfactant-like peptides, for performing such a "pulling-down" function. One or more of three such peptides (L6KD, L6K2, DKL6 were fused to the carboxyl termini of model proteins including Aspergillus fumigatus amadoriase II (AMA, all three peptides were used, Bacillus subtilis lipase A (LipA, only L6KD was used, hereinafter the same, Bacillus pumilus xylosidase (XynB, and green fluorescent protein (GFP, and expressed in E. coli. All fusions were found to predominantly accumulate in the insoluble fractions, with specific activities ranging from 25% to 92% of the native counterparts. Transmission electron microscopic (TEM and confocal fluorescence microscopic analyses confirmed the formation of protein aggregates in the cell. Furthermore, binding assays with amyloid-specific dyes (thioflavin T and Cong red to the AMA-L6KD aggregate and the TEM analysis of the aggregate following digestion with protease K suggested that the AMA-L6KD aggregate may contain structures reminiscent of amyloids, including a fibril-like structure core. Conclusions This study shows that the surfactant-like peptides L6KD and it derivatives can act as a pull-down handler for converting soluble proteins into active aggregates, much like 18A and ELK16. These peptide-mediated protein aggregations might have important implications for protein aggregation in

ORF-selector ESPRIT: a second generation library screen for soluble protein expression employing precise open reading frame selection.

Science.gov (United States)

An, Yingfeng; Yumerefendi, Hayretin; Mas, Philippe J; Chesneau, Alban; Hart, Darren J

2011-08-01

Here we present ORF-selector ESPRIT, a 9-fold enhanced version of our technology for screening incremental truncation libraries to identify soluble high yielding constructs of challenging proteins. Gene fragments are truncated at both termini to access internal domains and the resulting reading frame problem is addressed by an unbiased, intein-based open reading frame selection yielding only in-frame DNA inserts. This enriched library is then subcloned into a standard high-level expression plasmid where tens of thousands of constructs can be assayed in a two-step process using colony- and liquid-handling robots to isolate rare highly expressing clones useful for production of multi milligram quantities of purifiable proteins. The p85α protein was used to benchmark the system resulting in isolation of all known domains, either alone or in tandem. The human kinase IKK1 was then screened resulting in purification of a predicted internal domain. This strategy provides an integrated, facile route to produce soluble proteins from challenging and poorly understood target genes at quantities compatible with structural biology, screening applications and immunisation studies. The high genetic diversity that can be sampled opens the way to study more diverse systems including multisubunit complexes. Copyright © 2011 Elsevier Inc. All rights reserved.

DsSWEET17, a Tonoplast-Localized Sugar Transporter from Dianthus spiculifolius, Affects Sugar Metabolism and Confers Multiple Stress Tolerance in Arabidopsis

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Aimin Zhou

2018-05-01

Full Text Available Plant SWEETs (Sugars Will Eventually be Exported Transporters affect the growth of plants by regulating the transport of sugar from source to sink and its intracellular transport between different organelles. In this study, DsSWEET17 from Dianthus spiculifolius was identified and characterized. Real-time quantitative PCR analysis revealed that the expression of DsSWEET17 was affected by exogenous application of fructose and glucose as well as under salt, osmotic, and oxidation stress. Colocalization experiments showed that the DsSWEET17-GFP (green fluorescent protein fusion protein was localized to the FM4-64-labeled tonoplasts in Arabidopsis. Compared to the wild type, the transgenic Arabidopsis seedlings overexpressing DsSWEET17 had longer roots, greater fresh weight, and a faster root growth upon exogenous application of fructose. Furthermore, transgenic Arabidopsis seedlings had significantly higher fructose accumulation than was observed for the wild-type seedlings. The analysis of root length revealed that transgenic Arabidopsis had higher tolerance to salt, osmotic, and oxidative stresses. Taken together, our results suggest that DsSWEET17 may be a tonoplast sugar transporter, and its overexpression affects sugar metabolism and confers multiple stress tolerance in Arabidopsis.

DsSWEET17, a Tonoplast-Localized Sugar Transporter from Dianthus spiculifolius, Affects Sugar Metabolism and Confers Multiple Stress Tolerance in Arabidopsis.

Science.gov (United States)

Zhou, Aimin; Ma, Hongping; Feng, Shuang; Gong, Shufang; Wang, Jingang

2018-05-24

Plant SWEETs (Sugars Will Eventually be Exported Transporters) affect the growth of plants by regulating the transport of sugar from source to sink and its intracellular transport between different organelles. In this study, DsSWEET17 from Dianthus spiculifolius was identified and characterized. Real-time quantitative PCR analysis revealed that the expression of DsSWEET17 was affected by exogenous application of fructose and glucose as well as under salt, osmotic, and oxidation stress. Colocalization experiments showed that the DsSWEET17-GFP (green fluorescent protein) fusion protein was localized to the FM4-64-labeled tonoplasts in Arabidopsis . Compared to the wild type, the transgenic Arabidopsis seedlings overexpressing DsSWEET17 had longer roots, greater fresh weight, and a faster root growth upon exogenous application of fructose. Furthermore, transgenic Arabidopsis seedlings had significantly higher fructose accumulation than was observed for the wild-type seedlings. The analysis of root length revealed that transgenic Arabidopsis had higher tolerance to salt, osmotic, and oxidative stresses. Taken together, our results suggest that DsSWEET17 may be a tonoplast sugar transporter, and its overexpression affects sugar metabolism and confers multiple stress tolerance in Arabidopsis .

Differences between easy- and difficult-to-mill chickpea (Cicer arietinum L.) genotypes. Part III: free sugar and non-starch polysaccharide composition.

Science.gov (United States)

Wood, Jennifer A; Knights, Edmund J; Campbell, Grant M; Choct, Mingan

2014-05-01

Parts I and II of this series of papers identified several associations between the ease of milling and the chemical compositions of different chickpea seed fractions. Non-starch polysaccharides were implicated; hence, this study examines the free sugars and sugar residues. Difficult milling is associated with: (1) lower glucose and xylose residues (less cellulose and xyloglucans) and more arabinose, rhamnose and uronic acid in the seed coat, suggesting a more flexible seed coat that resists cracking and decortication; (2) a higher content of soluble and insoluble non-starch polysaccharide fractions in the cotyledon periphery, supporting a pectic polysaccharide mechanism comprising arabinogalacturonan, homogalacturonan, rhamnogalalcturonan, and glucuronan backbone structures; (3) higher glucose and mannose residues in the cotyledon periphery, supporting a lectin-mediated mechanism of adhesion; and (4) higher arabinose and glucose residues in the cotyledon periphery, supporting a mechanism involving arabinogalactan-proteins. This series has shown that the chemical composition of chickpea does vary in ways that are consistent with physical explanations of how seed structure and properties relate to milling behaviour. Seed coat strength and flexibility, pectic polysaccharide binding, lectins and arabinogalactan-proteins have been implicated. Increased understanding in these mechanisms will allow breeding programmes to optimise milling performance in new cultivars. © 2013 Society of Chemical Industry.

Evaluation of soluble solids content and pH of ice cream treated with gamma radiation

Energy Technology Data Exchange (ETDEWEB)

Rogovschi, V.D.; Nunes, T.C.F.; Fabbri, A.D.T.; Sagretti, J.M.; Sabato, S.F. [Instituto de Pesquisas Energeticas e Nucleares (IPEN/CNEN-SP), Sao Paulo, SP (Brazil)

2011-07-01

The ice cream is considered an aerated suspension of fat and ice crystals in a concentrated sugar solution containing hydrocolloids, proteins and casein micelles. Only in Brazil, in the year 2010, it was produced 1120 million liter of ice cream and due to high demand by the consumers, this is considered the most important product of the dairy industry. The objective of this work is to evaluate the soluble solids content (SSC) and the hydrogenionic potential (pH) of vanilla ice cream conditioned in isothermal boxes irradiated with 3.0 and 5.0 kGy in the Multipurpose Irradiator of {sup 60}Co located at IPEN - CNEN/SP. It can be concluded that the treatment of ice cream with gamma radiation did not cause changes in the analyzed parameters. . (author)

Evaluation of soluble solids content and pH of ice cream treated with gamma radiation

International Nuclear Information System (INIS)

Rogovschi, V.D.; Nunes, T.C.F.; Fabbri, A.D.T.; Sagretti, J.M.; Sabato, S.F.

2011-01-01

The ice cream is considered an aerated suspension of fat and ice crystals in a concentrated sugar solution containing hydrocolloids, proteins and casein micelles. Only in Brazil, in the year 2010, it was produced 1120 million liter of ice cream and due to high demand by the consumers, this is considered the most important product of the dairy industry. The objective of this work is to evaluate the soluble solids content (SSC) and the hydrogenionic potential (pH) of vanilla ice cream conditioned in isothermal boxes irradiated with 3.0 and 5.0 kGy in the Multipurpose Irradiator of 60 Co located at IPEN - CNEN/SP. It can be concluded that the treatment of ice cream with gamma radiation did not cause changes in the analyzed parameters. . (author)

Sugar ester surfactants: enzymatic synthesis and applications in food industry.

Science.gov (United States)

Neta, Nair S; Teixeira, José A; Rodrigues, Lígia R

2015-01-01

Sugar esters are non-ionic surfactants that can be synthesized in a single enzymatic reaction step using lipases. The stability and efficiency of lipases under unusual conditions and using non-conventional media can be significantly improved through immobilization and protein engineering. Also, the development of de novo enzymes has seen a significant increase lately under the scope of the new field of synthetic biology. Depending on the esterification degree and the nature of fatty acid and/or sugar, a range of sugar esters can be synthesized. Due to their surface activity and emulsifying capacity, sugar esters are promising for applications in food industry.

Sugar signalling and gene expression in relation to carbohydrate ...

Indian Academy of Sciences (India)

Sucrose is required for plant growth and development. The sugar status of plant cells is sensed by sensor proteins. The signal generated by signal transduction cascades, which could involve mitogen-activated protein kinases, protein phosphatases, Ca2+ and calmodulins, results in appropriate gene expression. A variety of ...

Endurance training blocks uncoupling protein 1 up-regulation in brown adipose tissue while increasing uncoupling protein 3 in the muscle tissue of rats fed with a high-sugar diet.

Science.gov (United States)

de Queiroz, Karina Barbosa; Rodovalho, Gisele Vieira; Guimarães, Juliana Bohnen; de Lima, Daniel Carvalho; Coimbra, Cândido Celso; Evangelista, Elísio Alberto; Guerra-Sá, Renata

2012-09-01

The mitochondrial uncoupling proteins (UCPs) of interscapular brown adipose tissue (iBAT) and of muscles play important roles in energy balance. For instance, the expression of UCP1 and UCP3 are modulated by free fatty acid gradients induced by high-sugar diets and acute exercise that is dependent on sympathetic stimulation. However, the effects of endurance training in animals fed with high-sugar diets are unknown. This study aims to evaluate the long-term effects of diet and exercise on UCP1 and UCP3 levels and energy balance efficiency. Rats fed with standard or high-sugar (HSD) diets were simultaneously subjected to running training over an 8-week period. After the training period, the rats were decapitated, and the iBAT and gastrocnemius muscle tissues were removed for evaluation of the β₃-receptor, Ucp1, and Ucp3 mRNA and protein expression, which were analyzed by quantitative reverse transcriptase polymerase chain reaction and Western blot, respectively. Groups fed with an HSD displayed a higher adiposity index and iBAT weight (P < .05), whereas exhibited an up-regulation of Ucp1 mRNA and protein levels (P < .05). Training increased β₃-receptor mRNA in iBAT and reduced the Ucp3 mRNA in muscle tissues. In association with an HSD, training restored the increasing β₃-receptor mRNA and greatly up-regulated the levels of Ucp3 mRNA. Therefore, training blocked the HSD-induced up-regulation of UCP1 expression in iBAT, whereas it up-regulated the expression of Ucp3 mRNA in muscle. These results suggest that training enhances the relationship between Ucp1/Ucp3 mRNA levels, which could result in higher energy efficiency, but not when HSD-induced elevated sympathetic activity is maintained. Copyright © 2012. Published by Elsevier Inc.

Water-Soluble Chlorophyll Protein (WSCP) Stably Binds Two or Four Chlorophylls.

Science.gov (United States)

Palm, Daniel M; Agostini, Alessandro; Tenzer, Stefan; Gloeckle, Barbara M; Werwie, Mara; Carbonera, Donatella; Paulsen, Harald

2017-03-28

Water-soluble chlorophyll proteins (WSCPs) of class IIa from Brassicaceae form tetrameric complexes containing one chlorophyll (Chl) per apoprotein but no carotenoids. The complexes are remarkably stable toward dissociation and protein denaturation even at 100 °C and extreme pH values, and the Chls are partially protected against photooxidation. There are several hypotheses that explain the biological role of WSCPs, one of them proposing that they function as a scavenger of Chls set free upon plant senescence or pathogen attack. The biochemical properties of WSCP described in this paper are consistent with the protein acting as an efficient and flexible Chl scavenger. At limiting Chl concentrations, the recombinant WSCP apoprotein binds substoichiometric amounts of Chl (two Chls per tetramer) to form complexes that are as stable toward thermal dissociation, denaturation, and photodamage as the fully pigmented ones. If more Chl is added, these two-Chl complexes can bind another two Chls to reach the fully pigmented state. The protection of WSCP Chls against photodamage has been attributed to the apoprotein serving as a diffusion barrier for oxygen, preventing its access to triplet excited Chls and, thus, the formation of singlet oxygen. By contrast, the sequential binding of Chls by WSCP suggests a partially open or at least flexible structure, raising the question of how WSCP photoprotects its Chls without the help of carotenoids.

A lactobacillus rhamnosus GG-derived soluble protein, p40, stimulates ligand release from intestinal epithelial cells to transactivate epidermal growth factor receptor

Science.gov (United States)

Protein p40, a Lactobacillus rhamnosus GG (LGG)-derived soluble protein, ameliorates intestinal injury and colitis, reduces apoptosis and preserves barrier function by activation of EGF receptor (EGFR) in intestinal epithelial cells. The aim of this study was to determine the mechanisms by which p40...

Effects of Bacterial Inoculants and Absorbents on Fermentation Properties and Chemical Composition of Fresh Sugar Beet Pulp Silage Using Laboratory silos

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Saeid Seidali Dolat-Abad

2016-04-01

Full Text Available Introduction Ensiling is one of the common preserving methods for forage or other organic materials. In this method, organic matters were preserved by proving an aerobic condition and then by reducing the pH with increasing acids production (mainly lactic acid. Some circumstances like enough soluble carbohydrates, low buffering capacity and appropriate dry matter concentration are needed in ensilages for an ideal silage production. Seepage production during ensiling is one of the most problems especially when high moisture materials (like fresh beet sugar pulp are ensiled. Silage seepage can pollute the environment and make loses in nutrients like soluble carbohydrates, protein, organic acids and etc. Moreover, lactic acid bacteria inoculants (Mainly consist of Lactobacillus plantarum have been widely used for improving fermentation pattern in ensilages. These external provided bacteria usually enhance lactic acid production in silage and then accelerate the falling of pH values in silages. Rapid decrease in pH can inhibit non-beneficial bacteria from activity which finally preserves nutrients from un-necessary fermentation or oxidation. The aim of this study was to investigate the interactive effects of lactic acid bacteria inoculants and some absorbents (straw and pith on chemical properties and fermentation profile of wet sugar beet pulp silage. Materials and Methods In the first experiment, fresh wet sugar beet pulp was treated with 5% straw or 5% pith in order to investigate the effects of these absorbents on chemical composition, fermentation characteristics and effluent production during ensiling period. In the second experiment, fresh wet sugar beet pulp was treated with a commercial lactic acid bacteria inoculants (Ecosyle and/or 5% pith in order to investing the main and interaction effects of the bacterial bacteria inoculants and the best absorbents from the experiment 1. In both experiments, triplicate samples were prepared for each

Effects of Mg on C and N Metabolism of Soybean at Different Nitrogen Supplying Levels

Institute of Scientific and Technical Information of China (English)

LANG Man; LIU Yuanying; PENG Xianlong; ZHANG Wenzhao

2006-01-01

A pot experiment was conducted to study the effects of magnesium on carbon and nitrogen metabolism of soybean at different nitrogen supplying levels. The results showed that the effects of magnesium at low nitrogen rate on N content, soluble protein, soluble sugar contents were not alike at different growth stage, although nodule dry weights raised, the yield and protein content of seeds decreased, however, the oil content was improved.The application of magnesium at medium and high nitrogen supplying levels promoted the uptake of N effectively,increased the soluble protein and soluble sugar contents, but the nodule dry weights of application magnesium at medium nitrogen supplying level decreased and the yield increased only a little despite the improved quality.Application of magnesium at high nitrogen supplying level raised nodule dry weights and soybean yield significantly, the quality of seeds was also improved.

Enzymatic gelation of sugar beet pectin in food products

DEFF Research Database (Denmark)

Bergsøe, Merete Norsker; Jensen, Mette; Adler-Nissen, Jens

2000-01-01

Sugar beet pectin is a food ingredient with specific functional properties. It may form gels by an oxidative cross-linking of ferulic acid. In the present study, the gel forming properties of three oxidative enzymes were examined in different food relevant conditions. The enzymes chosen were two...... laccases and one peroxidase. The textural properties of the produced gels were measured on a texture analyser. The influence of sugar, salt and protein were analysed. Finally, the enzymatic gelation was studied in three food products with added sugar beet pectin. These were black currant juice, milk...... and chopped heat-treated meat emulsion. The addition of salt resulted in softer, less stiff and chewy, and less adhesive gels. Generally speaking, sugar addition increased the hardness but at high concentration the gels were very brittle. However, Young's modulus was lower in gels containing sugar than...

Fermentation of rapeseed meal, sunflower meal and faba beans in combination with wheat bran increases solubility of protein and phosphorus

DEFF Research Database (Denmark)

Poulsen, Hanne Damgaard; Blaabjerg, Karoline

2017-01-01

BACKGROUND To increase self-supply of protein and phosphorus (P) in European pig and poultry diets and reduce nitrogen (N) and P excretion, attention is directed to approaches increasing protein and P digestibility of rapeseed, sunflower and faba beans. Wheat bran is rich in enzymes degrading...... and solubilizing protein and phytate. Herein, solubilization of protein, N and P was investigated when increasing ratios of wheat bran were fermented with rapeseed meal (RSM), sunflower meal (SFM), faba beans (FB) or a combination of these (RSM/SFM/FB). RESULTS Protein, N and P solubility was greater, for all...

Profiling of the Contents of Amino Acids, Water-Soluble Vitamins, Minerals, Sugars and Organic Acids in Turkish Hazelnut Varieties

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Taş Neslihan Göncüoğlu

2018-09-01

Full Text Available Proximate composition, profiles of amino acids, sugars, organic acids, vitamins and minerals of fourteen Turkish hazelnut varieties harvested in 2013 and 2014 were investigated. Glutamic acid, arginine and aspartic acid were the most predominant amino acids, representing of about 50% of hazelnut protein. Individual amino acid profiles showed significant differences depending upon the harvest year (p<0.05. Concentration of sucrose was the highest followed by fructose, glucose, stachyose, raffinose and myo-inositol, respectively. Phytic acid was predominant organic acid in all varieties, followed by malic acid. Independent of the variety, hazelnuts were rich in pantothenic acid, nicotinic acid, pyridoxal, biotin, thiamine, nicotinamide. Pantothenic and nicotinic acid were significantly higher in most of the varieties in harvest year 2014. Potassium was the most predominant mineral, followed by magnesium, calcium, sodium, manganese, zinc, iron and copper, respectively.

Co-expression of sulphydryl oxidase and protein disulphide isomerase in Escherichia coli allows for production of soluble CRM197

CSIR Research Space (South Africa)

Roth, Robyn L

2017-04-01

Full Text Available The aim of this article is to investigate the production of soluble cross-reacting material 197 (CRM(sub197)) in Escherichia coli, a safe and effective T-cell-dependent protein carrier for polysaccharides used in the manufacture and application...

Pronounced Phenotypic Changes in Transgenic Tobacco Plants Overexpressing Sucrose Synthase May Reveal a Novel Sugar Signaling Pathway

Science.gov (United States)

Nguyen, Quynh Anh; Luan, Sheng; Wi, Seung G.; Bae, Hanhong; Lee, Dae-Seok; Bae, Hyeun-Jong

2016-01-01

Soluble sugars not only serve as nutrients, but also act as signals for plant growth and development, but how sugar signals are perceived and translated into physiological responses in plants remains unclear. We manipulated sugar levels in transgenic plants by overexpressing sucrose synthase (SuSy), which is a key enzyme believed to have reversible sucrose synthesis and sucrose degradation functions. The ectopically expressed SuSy protein exhibited sucrose-degrading activity, which may change the flux of sucrose demand from photosynthetic to non-photosynthetic cells, and trigger an unknown sucrose signaling pathway that lead to increased sucrose content in the transgenic plants. An experiment on the transition from heterotrophic to autotrophic growth demonstrated the existence of a novel sucrose signaling pathway, which stimulated photosynthesis, and enhanced photosynthetic synthesis of sucrose, which was the direct cause or the sucrose increase. In addition, a light/dark time treatment experiment, using different day length ranges for photosynthesis/respiration showed the carbohydrate pattern within a 24-h day and consolidated the role of sucrose signaling pathway as a way to maintain sucrose demand, and indicated the relationships between increased sucrose and upregulation of genes controlling development of the shoot apical meristem (SAM). As a result, transgenic plants featured a higher biomass and a shorter time required to switch to reproduction compared to those of control plants, indicating altered phylotaxis and more rapid advancement of developmental stages in the transgenic plants. PMID:26793204

Sweet taste in apple: the role of sorbitol, individual sugars, organic acids and volatile compounds

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Aprea, Eugenio; Charles, Mathilde; Endrizzi, Isabella; Laura Corollaro, Maria; Betta, Emanuela; Biasioli, Franco; Gasperi, Flavia

2017-03-01

Sweetness is one of the main drivers of consumer preference, and thus is given high priority in apple breeding programmes. Due to the complexity of sweetness evaluation, soluble solid content (SSC) is commonly used as an estimation of this trait. Nevertheless, it has been demonstrated that SSC and sweet taste are poorly correlated. Though individual sugar content may vary greatly between and within apple cultivars, no previous study has tried to investigate the relationship between the amount of individual sugars, or ratios of these, and apple sweetness. In this work, we quantified the major sugars (sucrose, glucose, fructose, xylose) and sorbitol and explored their influence on perceived sweetness in apple; we also related this to malic acid content, SSC and volatile compounds. Our data confirmed that the correlation between sweetness and SSC is weak. We found that sorbitol content correlates (similarly to SSC) with perceived sweetness better than any other single sugar or total sugar content. The single sugars show no differentiable importance in determining apple sweetness. Our predictive model based on partial least squares regression shows that after sorbitol and SSC, the most important contribution to apple sweetness is provided by several volatile compounds, mainly esters and farnesene.

Decreased UV light resistance of spores of Bacillus subtilis strains deficient in pyrimidine dimer repair and small, acid-soluble spore proteins

International Nuclear Information System (INIS)

Setlow, B.; Setlow, P.

1988-01-01

Loss of small, acid-soluble spore protein alpha reduced spore UV resistance 30- to 50-fold in Bacillus subtilis strains deficient in pyrimidine dimer repair, but gave only a 5- to 8-fold reduction in UV resistance in repair-proficient strains. However, both repair-proficient and -deficient spores lacking this protein had identical heat and gamma-radiation resistance

Studies on soluble carbohydrates in yellow lupin (Lupinus luteus L. seeds of different age

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Kazimierz Zalewski

2011-01-01

Full Text Available Yellow lupin seeds cv. Juno were stored under laboratory conditions for 2 month, 4, 6 and 8 years. Eighteen soluble carbohydrates were identified in embryonic axes and cotyledons of different age seeds. The concentration of soluble carbohydrates in analysed seeds ranged from 25 to 34% of dry mass. Axes contained more carbohydrates than cotyledons. Stachyose dominated in axes, and verbascose - in cotyledons. Other detected galactosides were: galactinol, galactosyl pinitols and galactosyl chiro-inositols (fagopyritols, but their content was several-fold lower than that of RFOs (in both axes and cotyledons tissues. The concentration of soluble carbohydrates indicated, that sucrose to RFOs mass ratio, or other changes in sugars composition are not indicators of seed storage.

СHIPS FROM SUGAR BEET

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G. O. Magomedov

2014-01-01

Full Text Available Summary. Priority social problem in the Russian Federation is to provide diverse populations rational healthy diet, taking into account their traditions and economic status. Solving this problem requires the development of processing industries of agriculture on the basis of the improvement of existing and creation of new energy-saving environmentally friendly technologies that can provide deep, if possible without waste, recycling of raw materials. Therefore, the aim of research was the development of technology for production of sugar beet chips. Technology is as follows: sugar beet supplied into the washing machine to remove dirt from its surface. Washed roots inspect on conveyor belts. Next pure sugar beets sent to steam-heat treatment for cleaning the skin. After the beets is subjected to cutting by combining this process with a treatment with an aqueous solution of citric acid. Then he sent for the drying process is completed upon reaching a product of moisture content of 4-5 %. Drying chips feature is that under the high temperature reaction proceeds melanoidins between proteins and sugars present in sugar beet. As a result, the product obtained has the following characteristics: gold-yellow color; absence of a characteristic odor of sugar beet; pleasant sour taste; humidity of 4-5%. Thus, the new technology is relevant, because now the chips are one of the most popular products, ready to eat. A beet chips are rich in dietary fiber (pectin, hemicellulose and cellulose - 4-5 % minerals - macroelements (potassium, sodium, magnesium, calcium, phosphorus, trace elements (iron, zinc, copper, manganese - 0.5-0.6 %, and are the product of a functional food.

Rare sugar D-allose strongly induces thioredoxin-interacting protein and inhibits osteoclast differentiation in Raw264 cells.

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Yamada, Kana; Noguchi, Chisato; Kamitori, Kazuyo; Dong, Youyi; Hirata, Yuko; Hossain, Mohammad A; Tsukamoto, Ikuko; Tokuda, Masaaki; Yamaguchi, Fuminori

2012-02-01

Oxidative stress modulates the osteoclast differentiation via redox systems, and thioredoxin 1 (Trx) promotes the osteoclast formation by regulating the activity of transcription factors. The function of Trx is known to be regulated by its binding partner, thioredoxin-interacting protein (TXNIP). We previously reported that the expression of TXNIP gene is strongly induced by a rare sugar D-allose. In this study, we tested the hypothesis that D-allose could inhibit the osteoclast differentiation by regulating the Trx function. We used a murine Raw264 cell line that differentiates to the osteoclast by the receptor activator of nuclear factor-κB ligand (RANKL) treatment. The effect of sugars was evaluated by tartrate-resistant acid phosphatase staining. The expression and localization of TXNIP and Trx protein were examined by Western blotting and immunohistochemisty. The activity of the nuclear factor-κB, nuclear factor of activated T cells, and activator protein 1 transcription factors was measured by the luciferase reporter assay. The addition of D-allose (25 mmol/L) inhibited the osteoclast differentiation down to 9.53% ± 1.27% of a receptor activator of nuclear factor-κB ligand-only treatment. During the osteoclast differentiation, a significant increase of TNXIP was observed by D-allose treatment. The immunohistochemical analysis showed that both Trx and TXNIP existed in the nucleus in preosteoclasts and osteoclasts. Overexpression of TXNIP by plasmid transfection also inhibited the osteoclast formation, indicating the functional importance of TXNIP for the osteoclast differentiation. Transcriptional activity of the activator protein 1, nuclear factor-κB, and nuclear factor of activated T cells, known to be modulated by Trx, were inhibited by D-allose. In conclusion, our data indicate that D-allose is a strong inhibitor of the osteoclast differentiation, and this effect could be caused by TXNIP induction and a resulting inhibition of the Trx function

Elevated atmospheric CO2 affected photosynthetic products in wheat seedlings and biological activity in rhizosphere soil under cadmium stress.

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Jia, Xia; Liu, Tuo; Zhao, Yonghua; He, Yunhua; Yang, Mingyan

2016-01-01

The objective of this study was to investigate the effects of elevated CO2 (700 ± 23 μmol mol(-1)) on photosynthetic products in wheat seedlings and on organic compounds and biological activity in rhizosphere soil under cadmium (Cd) stress. Elevated CO2 was associated with decreased quantities of reducing sugars, starch, and soluble amino acids, and with increased quantities of soluble sugars, total sugars, and soluble proteins in wheat seedlings under Cd stress. The contents of total soluble sugars, total free amino acids, total soluble phenolic acids, and total organic acids in the rhizosphere soil under Cd stress were improved by elevated CO2. Compared to Cd stress alone, the activity of amylase, phenol oxidase, urease, L-asparaginase, β-glucosidase, neutral phosphatase, and fluorescein diacetate increased under elevated CO2 in combination with Cd stress; only cellulase activity decreased. Bacterial abundance in rhizosphere soil was stimulated by elevated CO2 at low Cd concentrations (1.31-5.31 mg Cd kg(-1) dry soil). Actinomycetes, total microbial abundance, and fungi decreased under the combined conditions at 5.31-10.31 mg Cd kg(-1) dry soil. In conclusion, increased production of soluble sugars, total sugars, and proteins in wheat seedlings under elevated CO2 + Cd stress led to greater quantities of organic compounds in the rhizosphere soil relative to seedlings grown under Cd stress only. Elevated CO2 concentrations could moderate the effects of heavy metal pollution on enzyme activity and microorganism abundance in rhizosphere soils, thus improving soil fertility and the microecological rhizosphere environment of wheat under Cd stress.

To drink or grasp? How bullet ants ( Paraponera clavata) differentiate between sugars and proteins in liquids

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Jandt, Jennifer; Larson, Hannah K.; Tellez, Peter; McGlynn, Terrence P.

2013-12-01

Flexibility in behavior can increase the likelihood that a forager may respond optimally in a fluctuating environment. Nevertheless, physiological or neuronal constraints may result in suboptimal responses to stimuli. We observed foraging workers of the giant tropical ant (also referred to as the "bullet ant"), Paraponera clavata, as they reacted to liquid solutions with varying concentrations of sugar and protein. We show that when protein/sucrose concentration is high, many bullet ants will often try to grasp at the droplet, rather than gather it by drinking. Because P. clavata actively hunt for prey, fixed action patterns and rapid responses to protein may be adaptively important, regardless of the medium in which it is presented. We conclude that, in P. clavata, food-handling decisions are made in response to the nutrient content of the food rather than the texture of the food. Further, we suggest that colonies that maintain a mixture of individuals with consistent fixed or flexible behavioral responses to food-handling decisions may be better adapted to fluctuating environmental conditions, and we propose future studies that could address this.

Responses to sugar and sugar receptor gene expression in different social roles of the honeybee (Apis mellifera).

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Değirmenci, Laura; Thamm, Markus; Scheiner, Ricarda

2018-04-01

Honeybees (Apis mellifera) are well-known for their sophisticated division of labor with each bee performing sequentially a series of social tasks. Colony organization is largely based on age-dependent division of labor. While bees perform several tasks inside the hive such as caring for brood ("nursing"), cleaning or sealing brood cells or producing honey, older bees leave to colony to collect pollen (proteins) and nectar (carbohydrates) as foragers. The most pronounced behavioral transition occurs when nurse bees become foragers. For both social roles, the detection and evaluation of sugars is decisive for optimal task performance. Nurse bees rely on their gustatory senses to prepare brood food, while foragers evaluate a nectar source before starting to collect food from it. To test whether social organization is related to differential sensing of sugars we compared the taste of nurse bees and foragers for different sugars. Searching for molecular correlates for differences in sugar perception, we further quantified expression of gustatory receptor genes in both behavioral groups. Our results demonstrate that nurse bees and foragers perceive and evaluate different sugars differently. Both groups, however, prefer sucrose over fructose. At least part of the taste differences between social roles could be related to a differential expression of taste receptors in the antennae and brain. Our results suggest that differential expression of sugar receptor genes might be involved in regulating division of labor through nutrition-related signaling pathways. Copyright © 2017 Elsevier Ltd. All rights reserved.

Stabilization of apoglobin by low temperature increases yield of soluble recombinant hemoglobin in Escherichia coli.

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Weickert, M J; Pagratis, M; Curry, S R; Blackmore, R

1997-01-01

Accumulation of soluble recombinant hemoglobin (rHb1.1) in Escherichia coli requires proper protein folding, prosthetic group (heme) addition, and subunit assembly. This served as a new model system for the study of the effects of temperature, protein synthesis rates, and protein accumulation rates on protein solubility in E. coli. Fermentation expression of rHb1.1 at 30 degrees C from cultures containing a medium or high globin gene dosage (pBR-based or pUC-based plasmids with rHb1.1 genes under the control of the tac promoter) was compared. A medium gene dosage resulted in rHb1.1 accumulating to approximately 7% of the soluble cell protein, of which 78% was soluble. A high globin gene dosage resulted in a > or = 3-fold increase in total globin to 23 to 24% of the soluble cell protein, but 70% was insoluble. Accumulation of insoluble rHb1.1 began immediately upon induction. The proportion of rHb1.1 from the high globin gene dosage that accumulated as insoluble globin was affected by reducing (i) the inducer concentration and (ii) the temperature. Reducing the inducer concentration reduced globin synthesis up to eightfold but increased the proportion of soluble rHb1.1 to 93%. In contrast, total globin protein synthesis was barely affected by reducing the temperature from 30 to 26 degrees C, while soluble globin accumulation increased > 2-fold to approximately 15% of the soluble cell protein. The contrast between the effects of reducing rates of protein synthesis and accumulation and those of reducing temperature suggests that lower temperature stabilizes one or more folding intermediates. We propose a simplified physical model which integrates protein synthesis, folding, and heme association. This model shows that temperature-dependent apoglobin stability is the most critical factor in soluble rHb1.1 accumulation. PMID:9361418

Effect of electrical stunning frequency on meat quality, plasma parameters, and protein solubility of broilers.

Science.gov (United States)

Huang, J C; Yang, J; Zhang, B H; Huang, M; Chen, K J; Xu, X L; Zhou, G H

2017-08-01

This study was designed to compare the effects of different stunning frequencies of pulsed direct current on meat quality of broilers. This was achieved by investigating plasma parameters, blood loss, carcass damage, meat water-holding capacity, meat color, meat shear value, muscle pH, and protein solubility. A total of 400 broilers was divided into 5 treatment groups and stunned with 500, 600, 700, 800, and 900 Hz at 15 V for 10 seconds. Blood samples were collected immediately after cutting the neck. Pectoralis major muscles were removed from the carcass after chilling and placed in ice. Breast muscle pH and meat color were determined at both 2 and 24 h postmortem. Drip loss, cooking loss, pressing loss, and cooked breast meat-shear values were determined at 24 h postmortem. Treatment at 500 and 900 Hz significantly increased (P meat color were not affected by stunning frequency. In the 500 and 900 Hz groups, the protein solubility and shear force values were significantly lower (P < 0.05) and drip loss was significantly higher (P < 0.05) than in the 700 Hz group. This study indicates that the waveform of the pulsed direct current is acceptable for stunning broilers at a stunning frequency of 700 Hz. © 2017 Poultry Science Association Inc.

Efficient expression of a soluble lipid transfer protein (LTP) of Platanus orientalis using short peptide tags and structural comparison with the natural form.

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Salari, Farhad; Vahedi, Fatemeh; Chamani, Jamshidkhan; Varasteh, Abdolreza; Ketabdar, Hanieh; Sankian, Mojtaba

2015-01-01

Successful recombinant allergen-based immunotherapy has drawn a great deal of attention to use recombinant allergens for new therapeutic and/or diagnostic strategies. The Escherichia coli expression system is frequently used to produce recombinant allergens; however, protein expression in E. coli often results in inclusion bodies. Here, we focused on the expression of two recombinant soluble forms of Pla or 3 using solubility-enhancing peptide tags, human immune deficiency virus type 1 transactivator of transcription core domain and poly-arginine-lysine: rTAT-Pla or 3 and rPoly-Arg-Lys-Pla or 3. Structural characteristics and IgE reactivity of purified recombinant proteins were compared with natural Pla or 3 (nPla or 3) isolated from Platanus orientalis using circular dichroism spectra, fluorescence spectroscopy, and immunoblotting. Likewise, intrinsic viscosity and Stokes radius of the natural and recombinant Pla or 3 allergens were determined to analyze structural compactness in aqueous media. The results indicate high-level solubility and efficient expression of the fusion proteins (rTAT-Pla or 3 and rPoly-Arg-Lys-Pla or 3) compared with the wild-type recombinant. Furthermore, the similar structural characteristics and IgE-binding activities of the fusion proteins to nPla or 3 provide a promising tool for allergy diagnosis and treatment. © 2014 International Union of Biochemistry and Molecular Biology, Inc.

Effects of photoperiod, growth temperature and cold acclimatisation on glucosinolates, sugars and fatty acids in kale.

Science.gov (United States)

Steindal, Anne Linn Hykkerud; Rødven, Rolf; Hansen, Espen; Mølmann, Jørgen

2015-05-01

Curly kale is a robust, cold tolerant plant with a high content of health-promoting compounds, grown at a range of latitudes. To assess the effects of temperature, photoperiod and cold acclimatisation on levels of glucosinolates, fatty acids and soluble sugars in kale, an experiment was set up under controlled conditions. Treatments consisted of combinations of the temperatures 15/9 or 21/15 °C, and photoperiods of 12 or 24h, followed by a cold acclimatisation period. Levels of glucosinolates and fatty acid types in leaves were affected by growth conditions and cold acclimatisation, being generally highest before acclimatisation. The effects of growth temperature and photoperiod on freezing tolerance were most pronounced in plants grown without cold acclimatisation. The results indicate that cold acclimatisation can increase the content of soluble sugar and can thereby improve the taste, whilst the content of unsaturated fatty and glucosinolates acids may decrease. Copyright © 2014 Elsevier Ltd. All rights reserved.

Studying of Salinity Stress Effect on Germination, Proline, Sugar, Protein, Lipid and Chlorophyll Content in Purslane (Portulaca oleracea L. Leaves

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Parvaneh Rahdari

2012-04-01

Full Text Available Approximately 30% of country regions are under influence of salt and one of the main important methods for increasing of hectare performance in salt soils, using of resistant plant against with salt such as medicinal annual plant of Purslane (Portulaca oleraceae L..Studying of the salt stress effect on germination and some physiological parameters in Purslane was carried out in completely random framework and with three replications and under controlled conditions. In this study, salinity was imposed by sodium chloride (NaCl and in five levels of framework included of Control (distilled water, 50, 100, 150 and 200 Mmol from NaCl. Studied physiological parameters included of proline, sugar, protein, lipid and chlorophyll in Purslane leaves.In relation with observed germination that increasing of salt level, germination rate was reduced that the least germination rate in 200 Mmol and the highest germination was observed in control which in statistical level 0.01 showed meaningful difference. Also, results showed an increasing of leaf proline and sugar that was synchronize with increasing of sodium chloride, this increasing in statistical level of P<0.01 was meaningful, while blank had the least rate and salt level 200 Mmol was observed. Synchronize with increasing of salinity level, lipid and protein concentration decreasing that was meaningful (P<0.01 which highest of leaf lipid and protein content in control level and least concentration was observed in 200 Mmol of salinity level. Also, salinity stress due to increasing of chlorophyll a and b levels in Purslane leaves which this increasing in leaf chlorophyll b concentrations in 0.01 statistical level was meaningful and the least level chlorophyll a and b in blank and the highest level of chlorophyll a and b by order in 150 and 200 Mmol of NaCl was observed. The results showed that plant for resistance against salinity increasing of proline, sugar and chlorophyll content in leaves are useful for

Anti-cariogenic properties of a water-soluble extract from cacao.

Science.gov (United States)

Ito, Kyoko; Nakamura, Yuko; Tokunaga, Takahisa; Iijima, Daisuke; Fukushima, Kazuo

2003-12-01

The addition of a water-soluble extract from cacao-extracted powder (CEPWS) to a cariogenic model food, a white chocolate-like diet that contains 35% sucrose, significantly reduced caries scores in SPF rats infected with Streptococcus sobrinus 6715, compared to control rats fed a white chocolate-like diet. CEPWS markedly inhibited water-insoluble glucan (WIG) synthesis through crude glucosyltransferases (GTFs) from Streptococcus sobrinus B13N in vitro. GTF-inhibitor(s) in CEPWS was prepared through three-step fractionation, and was termed CEPWS-BT, which is a high molecular weight (>10 kDa) heat-stable matrix of sugar, protein, and polyphenol. When the inhibitory effect of CEPWS-BT on glucan synthesis was examined using the purified GTF-I, GTF-T, and GTF-U enzymes from S. sobrinus B13N, significant reduction in GTF-I and GTF-T activity as a result of adding CEPWS-BT at low concentrations was observed. These results suggest that the addition of CEPWS to cariogenic food could be useful in controlling dental caries.

Sugar cane bagasse prehydrolysis using hot water

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D. Abril

2012-03-01

Full Text Available Results are presented on the hot water prehydrolysis of sugar cane bagasse for obtaining ethanol by fermentation. The experimental study consisted of the determination of the effect of temperature and time of prehydrolysis on the extraction of hemicelluloses, with the objective of selecting the best operating conditions that lead to increased yield of extraction with a low formation of inhibitors. The study, carried out in a pilot plant scale rotational digester, using a 3² experimental design at temperatures of 150-190ºC and times of 60-90 min, showed that it is possible to perform the hot water prehydrolysis process between 180-190ºC in times of 60-82 min, yielding concentrations of xylose > 35 g/L, furfural < 2.5 g/L, phenols from soluble lignin < 1.5 g/L, and concentrations < 3.0 g/L of hemicelluloses in the cellolignin residue. These parameters of temperature and prehydrolysis time could be used for the study of the later hydrolysis and fermentation stages of ethanol production from sugar cane bagasse.

Scoring function to predict solubility mutagenesis

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Deutsch Christopher

2010-10-01

Full Text Available Abstract Background Mutagenesis is commonly used to engineer proteins with desirable properties not present in the wild type (WT protein, such as increased or decreased stability, reactivity, or solubility. Experimentalists often have to choose a small subset of mutations from a large number of candidates to obtain the desired change, and computational techniques are invaluable to make the choices. While several such methods have been proposed to predict stability and reactivity mutagenesis, solubility has not received much attention. Results We use concepts from computational geometry to define a three body scoring function that predicts the change in protein solubility due to mutations. The scoring function captures both sequence and structure information. By exploring the literature, we have assembled a substantial database of 137 single- and multiple-point solubility mutations. Our database is the largest such collection with structural information known so far. We optimize the scoring function using linear programming (LP methods to derive its weights based on training. Starting with default values of 1, we find weights in the range [0,2] so that predictions of increase or decrease in solubility are optimized. We compare the LP method to the standard machine learning techniques of support vector machines (SVM and the Lasso. Using statistics for leave-one-out (LOO, 10-fold, and 3-fold cross validations (CV for training and prediction, we demonstrate that the LP method performs the best overall. For the LOOCV, the LP method has an overall accuracy of 81%. Availability Executables of programs, tables of weights, and datasets of mutants are available from the following web page: http://www.wsu.edu/~kbala/OptSolMut.html.

Impacto dos nutrientes N e K e de açúcares solúveis sobre populações de Diabrotica speciosa (Germar (Coleoptera, Chrysomelidae e Agrotis ipsilon (Hüfnagel (Lepidoptera, Noctuidae na cultura da batata, Solanum tuberosum L. (Solanaceae Impact of the nutrients N and K and soluble sugars on Diabrotica speciosa (Germar (Coleoptera, Chrysomelidae and Agrotis ipsilon (Hüfnagel (Lepidoptera, Noctuidae populations in potato crops, Solanum tuberosum L. (Solanaceae

Directory of Open Access Journals (Sweden)

Edson Henrique de Azeredo

2004-03-01

Full Text Available Foi estudada a ocorrência de Diabrotica speciosa (Germar, 1824 (Coleoptera, Chrysomelidae e de Agrotis ipsilon (Hüfnagel, 1767 (Lepidoptera, Noctuidae em plantas de batata, cultivares Achat e Monalisa, influenciadas por dosagens de nitrogênio e potássio, e teor mínimo de açúcares solúveis. Os seguintes parâmetros foram avaliados: concentração de nutrientes minerais e açúcar em folha verde, folha senescente, folha em abcisão, haste, tubérculo e planta total usando extratos de infusão em etanol 80%. A maior infestação por larvas de D. speciosa foi na cultivar Monalisa a 150 kg.ha-1 de N + K com 27,03% a PThe occurrence of Diabrotica speciosa (Germar, 1824 and Agrotis ipsilon (Hüfnagel, 1767 on the potato cultivars Achat and Monalisa, influenced by nitrogen and potassium dosage, and minimum theor of soluble sugars, was studied. The following parameters were evaluated: concentration of mineral nutrient and sugar in green leaf, senescent leaf, leaf in abscission, stem, tubercle and total plant using extracts of infusion in ethanol 80%. The largest infestation of D. speciosa larvae was on Monalisa cultivar at 150 kg.ha-1 of N + K with 27.03% at P< 0,05. It was observed that the effect of the dosage of N + K in the increment of the concentration of soluble sugars increased the damages in the tubercles and stems by A. ipsilon. The infestation by these species increased to 58.82% on the Monalisa cultivar, when the nitrogen dosage increased from zero to 150 kg.ha-1, in the absence of potassium. On the other hand, high dosage of K reduced the damages by A. ipsilon on Monalisa cultivar. However, it did not influence the storage of soluble sugar. The results indicated that in Achat cultivar the accumulated soluble sugar was reduced, probably sensibilized by elevation of potassic fertilization dosing, differing from Monalisa cultivar, in which the influence was by nitrogen dosing.

Overcoming the solubility limit with solubility-enhancement tags: successful applications in biomolecular NMR studies

International Nuclear Information System (INIS)

Zhou Pei; Wagner, Gerhard

2010-01-01

Although the rapid progress of NMR technology has significantly expanded the range of NMR-trackable systems, preparation of NMR-suitable samples that are highly soluble and stable remains a bottleneck for studies of many biological systems. The application of solubility-enhancement tags (SETs) has been highly effective in overcoming solubility and sample stability issues and has enabled structural studies of important biological systems previously deemed unapproachable by solution NMR techniques. In this review, we provide a brief survey of the development and successful applications of the SET strategy in biomolecular NMR. We also comment on the criteria for choosing optimal SETs, such as for differently charged target proteins, and recent new developments on NMR-invisible SETs.

Effect of urdbean leaf crinkle virus infection on total soluble protein and antioxidant enzymes in blackgram plants

International Nuclear Information System (INIS)

Ashfaq, M.; Mughal, S.M.; Khan, A.; Javed, N.; Sahi, S.T.; Shahid, M.

2010-01-01

Urdbean leaf crinkle virus (ULCV) is a common, wide spread, destructive and economically important disease causing systemic infection in blackgram (Vigna mungo (L.) Hepper), resulting in extreme crinkling, curling, puckering and rugosity of leaves, and yield reductions. Effect of viral infection was investigated on total soluble proteins and antioxidant enzymes activity in two genotypes viz., Mash-88-susceptible and CM-2002-resistant, at different growth stages under both the inoculated and un-inoculated conditions. ULCV infection resulted in significant increase in total soluble protein contents of the leaves in both genotypes. In healthy plant, super oxide dismutase (SOD), catalase (CAT) and peroxidase (PO) showed similar activity levels. In inoculated plants of Mash-88, SOD and PO activities decreased and increased non-significantly at all growth stages, respectively. The activities of PO and SOD increased and decreased significantly after 15 and 30 days of inoculation in resistant genotype, respectively. No significant changes in catalase (CAT) activity were detected in ULCV-infected leaves over the control. It was concluded that the super oxide dismutase and peroxidases might be associated with resistance/susceptibility to ULCV infection. (author)

Bee venom phospholipase A2 as a membrane-binding vector for cell surface display or internalization of soluble proteins.

Science.gov (United States)

Babon, Aurélie; Wurceldorf, Thibault; Almunia, Christine; Pichard, Sylvain; Chenal, Alexandre; Buhot, Cécile; Beaumelle, Bruno; Gillet, Daniel

2016-06-15

We showed that bee venom phospholipase A2 can be used as a membrane-binding vector to anchor to the surface of cells a soluble protein fused to its C-terminus. ZZ, a two-domain derivative of staphylococcal protein A capable of binding constant regions of antibodies was fused to the C-terminus of the phospholipase or to a mutant devoid of enzymatic activity. The fusion proteins bound to the surface of cells and could themselves bind IgGs. Their fate depended on the cell type to which they bound. On the A431 carcinoma cell line the proteins remained exposed on the cell surface. In contrast, on human dendritic cells the proteins were internalized into early endosomes. Copyright © 2015 Elsevier Ltd. All rights reserved.

Cytokine-like factor-1, a novel soluble protein, shares homology with members of the cytokine type I receptor family.

Science.gov (United States)

Elson, G C; Graber, P; Losberger, C; Herren, S; Gretener, D; Menoud, L N; Wells, T N; Kosco-Vilbois, M H; Gauchat, J F

1998-08-01

In this report we describe the identification, cloning, and expression pattern of human cytokine-like factor 1 (hCLF-1) and the identification and cloning of its murine homologue. They were identified from expressed sequence tags using amino acid sequences from conserved regions of the cytokine type I receptor family. Human CLF-1 and murine CLF-1 shared 96% amino acid identity and significant homology with many cytokine type I receptors. CLF-1 is a secreted protein, suggesting that it is either a soluble subunit within a cytokine receptor complex, like the soluble form of the IL-6R alpha-chain, or a subunit of a multimeric cytokine, e.g., IL-12 p40. The highest levels of hCLF-1 mRNA were observed in lymph node, spleen, thymus, appendix, placenta, stomach, bone marrow, and fetal lung, with constitutive expression of CLF-1 mRNA detected in a human kidney fibroblastic cell line. In fibroblast primary cell cultures, CLF-1 mRNA was up-regulated by TNF-alpha, IL-6, and IFN-gamma. Western blot analysis of recombinant forms of hCLF-1 showed that the protein has the tendency to form covalently linked di- and tetramers. These results suggest that CLF-1 is a novel soluble cytokine receptor subunit or part of a novel cytokine complex, possibly playing a regulatory role in the immune system and during fetal development.

Reduction in lipophilicity improved the solubility, plasma–protein binding, and permeability of tertiary sulfonamide RORc inverse agonists

Energy Technology Data Exchange (ETDEWEB)

Fauber, Benjamin P.; René, Olivier; de Leon Boenig, Gladys; Burton, Brenda; Deng, Yuzhong; Eidenschenk, Céline; Everett, Christine; Gobbi, Alberto; Hymowitz, Sarah G.; Johnson, Adam R.; La, Hank; Liimatta, Marya; Lockey, Peter; Norman, Maxine; Ouyang, Wenjun; Wang, Weiru; Wong, Harvey (Genentech); (Argenta)

2014-08-01

Using structure-based drug design principles, we identified opportunities to reduce the lipophilicity of our tertiary sulfonamide RORc inverse agonists. The new analogs possessed improved RORc cellular potencies with >77-fold selectivity for RORc over other nuclear receptors in our cell assay suite. The reduction in lipophilicity also led to an increased plasma–protein unbound fraction and improvements in cellular permeability and aqueous solubility.

Action of mercurials on activity of partially purified soluble protein kinase C from mice brain

International Nuclear Information System (INIS)

Inoue, Y.; Saijoh, K.; Sumino, K.

1988-01-01

The enzymatic activity of soluble protein kinase C from mice brain was inhibited by mercuric chloride (II) (HgCl 2 ) and organic mercurials, i.e. methyl mercury, phenyl mercury and p-chloromercuribenzoic acid (PCMB). The IC50 was 0.08 μM for HgCl 2 and about 1 μM for organic mercurials. Sulfhydryl blocking reagents such as 5.5'-dithiobis-2-nitrobenzoic acid (DTNB) and N-ethylmaleimide (NEM) were less potent but nevertheless inhibited the enzymic activity of protein kinase C. The Hill coefficients of HgCl 2 , DTNB and NEM were close to unity whereas the values for organic mercurials were 1.3 to 1.5. The inhibition was of a non-competitive type with respect to Hl histone. 3 H-PDBu binding activity was also inhibited by all of the reagents in a non-competitive manner. Mercurials apparently bind to sulfhydryl groups of protein kinase C to inhibit the enzymatic activity. (author)

Phenome data - High-sugar stress - DGBY | LSDB Archive [Life Science Database Archive metadata

Lifescience Database Archive (English)

Full Text Available switchLanguage; BLAST Search Image Search Home About Archive Update History Data ...s.zip File size: 90KB File name: Original file: High-sugar_stress.xls File URL: ftp://ftp.biosciencedbc.jp/archive/dgby/LATEST/Hi... Center for Protein Sequences） About This Database Database Description Download License Update History of Thi...s Database Site Policy | Contact Us Phenome data - High-sugar stress - DGBY | LSDB Archive ... ...List Contact us DGBY Phenome data - High-sugar stress Data detail Data name Phenome data - High-sugar stress

Design of sweet protein based sweeteners: hints from structure-function relationships.

Science.gov (United States)

Rega, Michele Fortunato; Di Monaco, Rossella; Leone, Serena; Donnarumma, Federica; Spadaccini, Roberta; Cavella, Silvana; Picone, Delia

2015-04-15

Sweet proteins represent a class of natural molecules, which are extremely interesting regarding their potential use as safe low-calories sweeteners for individuals who need to control sugar intake, such as obese or diabetic subjects. Punctual mutations of amino acid residues of MNEI, a single chain derivative of the natural sweet protein monellin, allow the modulation of its taste. In this study we present a structural and functional comparison between MNEI and a sweeter mutant Y65R, containing an extra positive charge on the protein surface, in conditions mimicking those of typical beverages. Y65R exhibits superior sweetness in all the experimental conditions tested, has a better solubility at mild acidic pH and preserves a significant thermal stability in a wide range of pH conditions, although slightly lower than MNEI. Our findings confirm the advantages of structure-guided protein engineering to design improved low-calorie sweeteners and excipients for food and pharmaceutical preparations. Copyright © 2014 Elsevier Ltd. All rights reserved.

Use of Pleurotus sajor-caju in upgrading green jute plants and jute sticks as ruminant feed

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Zinat Mahal

2013-01-01

Full Text Available In this study, superfluous jute plants and jute stick were converted into upgraded animal feed by solid state fermentation (SSF using a cellulolytic fungus, Pleurotus sajor-caju. Prior to fermentation, substrates were subjected to several pretreatments such as soaking with water overnight and alkali or lime pretreatment. SSF was carried out with 20 g of substrate in 100 ml conical flask and was incubated at 30C for 8 weeks. In all treatments, the highest amount of reducing sugar, soluble protein as well as the cellulolytic activities of three enzymes viz. cellobiase, carboxymethyl cellulase and avicelase were obtained at 6th week of fermentation. Compared to raw, unsoaked substrates, soaking treatment alone could produce 10% more soluble protein in both substrates whereas reducing sugar increment was 5% and 6% in jute sticks and jute plants, respectively. From all treatments, combination of soaking and lime treatment in green jute plants yielded higher value than jute sticks in terms of reducing sugar, soluble protein and enzymatic activity. The radiation doses at 20, 30 and 40 kGy appeared to have no effect on sugar and protein accretion. During eight weeks of fermentation, relatively higher cellobiase activity was found compared to that of carboxymethyl cellulase and avicelase. The present investigation indicates that fungal conversion with pretreatment can turn these lignocellulosic agro-wastes to a nutritionally enriched animal feed by increasing the crude protein and reducing sugar content. However, further research is necessary to develop strategies for industrial scale production to overcome the crisis of nutritionally improved animal feed.

Biochemical characterisation and dietary fibre analysis of sugar beet supplemented cookies

International Nuclear Information System (INIS)

Pasha, I.; Jahangir, M.F.; Akhter, S.; Manzoor, M.S.

2016-01-01

This study was planned to utilize sugar beet powder as a rich source of dietary fibre in cookies. Purposely, five treatments namely T1, T2, T3, T4 and T5 with 4%, 8%, 12%, 16% and 20% sugar beet powder addition in wheat flour were chosen to estimate fibre, antioxidant profiling and engineering properties of cookies. Results showed an increased content of all above mentioned parameters. With the increment in sugar beet powder addition in treatments, dietary fibre analysis have shown that total dietary fibre (TDF), insoluble dietary fibre (IDF) and soluble dietary fibre (SDF) have depicted increasing trend with maximum for T5 for all dietary fibre types. Significant results were obtained for in vitro antioxidant studies including total phenolic content (TPC) and DPPH that showed increasing trend with T1 0.6 mg GAE/g and maximum values for T5 with 2.0 mg GAE/g for TPC and for DPPH with T5 being maximum value of 1.7% and minimum for T1 with 1.3%. T5 treatment with 20% sugar beet gave best physicochemical results but disturbed sensory properties while T3 with 12% sugar beet powder showed good physicochemical and sensory characteristics. Therefore, T3 with 12% level is considered as the best source of dietary fibre in bakery products and can be considered as the prospective choice to address metabolic syndromes. (author)

Soluble CD163

DEFF Research Database (Denmark)

Møller, Holger J

2012-01-01

CD163 is an endocytic receptor for haptoglobin-hemoglobin complexes and is expressed solely on macrophages and monocytes. As a result of ectodomain shedding, the extracellular portion of CD163 circulates in blood as a soluble protein (sCD163) at 0.7-3.9 mg/l in healthy individuals. The function o...

Saccharification of recalcitrant biomass and integration options for lignocellulosic sugars from Catchlight Energy's sugar process (CLE Sugar).

Science.gov (United States)

Gao, Johnway; Anderson, Dwight; Levie, Benjamin

2013-01-28

Woody biomass is one of the most abundant biomass feedstocks, besides agriculture residuals in the United States. The sustainable harvest residuals and thinnings alone are estimated at about 75 million tons/year. These forest residuals and thinnings could produce the equivalent of 5 billion gallons of lignocellulosic ethanol annually. Softwood biomass is the most recalcitrant biomass in pretreatment before an enzymatic hydrolysis. To utilize the most recalcitrant lignocellulosic materials, an efficient, industrially scalable and cost effective pretreatment method is needed. Obtaining a high yield of sugar from recalcitrant biomass generally requires a high severity of pretreatment with aggressive chemistry, followed by extensive conditioning, and large doses of enzymes. Catchlight Energy's Sugar process, CLE Sugar, uses a low intensity, high throughput variation of bisulfite pulping to pretreat recalcitrant biomass, such as softwood forest residuals. By leveraging well-proven bisulfite technology and the rapid progress of enzyme suppliers, CLE Sugar can achieve a high yield of total biomass carbohydrate conversion to monomeric lignocellulosic sugars. For example, 85.8% of biomass carbohydrates are saccharified for un-debarked Loblolly pine chips (softwood), and 94.0% for debarked maple chips (hardwood). Furan compound formation was 1.29% of biomass feedstock for Loblolly pine and 1.10% for maple. At 17% solids hydrolysis of pretreated softwood, an enzyme dose of 0.075 g Sigma enzyme mixture/g dry pretreated (unwashed) biomass was needed to achieve 8.1% total sugar titer in the hydrolysate and an overall prehydrolysate liquor plus enzymatic hydrolysis conversion yield of 76.6%. At a much lower enzyme dosage of 0.044 g CTec2 enzyme product/g dry (unwashed) pretreated softwood, hydrolysis at 17% solids achieved 9.2% total sugar titer in the hydrolysate with an overall sugar yield of 85.0% in the combined prehydrolysate liquor and enzymatic hydrolysate. CLE Sugar has

High-level expression and purification of soluble recombinant FGF21 protein by SUMO fusion in Escherichia coli

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Huang Yadong

2010-02-01

Full Text Available Abstract Background Fibroblast growth factor 21 (FGF21 is a promising drug candidate to combat metabolic diseases. However, high-level expression and purification of recombinant FGF21 (rFGF21 in Escherichia coli (E. coli is difficult because rFGF21 forms inclusion bodies in the bacteria making it difficult to purify and obtain high concentrations of bioactive rFGF21. To overcome this problem, we fused the FGF21 with SUMO (Small ubiquitin-related modifier by polymerase chain reaction (PCR, and expressed the fused gene in E. coli BL21(DE3. Results By inducing with IPTG, SUMO-FGF21 was expressed at a high level. Its concentration reached 30% of total protein, and exceeded 95% of all soluble proteins. The fused protein was purified by DEAE sepharose FF and Ni-NTA affinity chromatography. Once cleaved by the SUMO protease, the purity of rFGF21 by high performance liquid chromatography (HPLC was shown to be higher than 96% with low endotoxin level (in vivo animal experiments showed that rFGF21 produced by using this method, could decrease the concentration of plasma glucose in diabetic rats by streptozotocin (STZ injection. Conclusions This study demonstrated that SUMO, when fused with FGF21, was able to promote its soluble expression of the latter in E. coli, making it more convenient to purify rFGF21 than previously. This may be a better method to produce rFGF21 for pharmaceutical research and development.

Sugar Sugar – don’t be misled / laat je niet misleiden

NARCIS (Netherlands)

Toebes, Brigit

2017-01-01

NRC Handelsblad’s Saturday 25 November issue contains an entry of eleven pages entirely devoted to sugar. It discusses a broad range of topics related to sugar, including the role of sugar throughout the centuries, sugar consumption in the Netherlands, the amount of sugar in bread, and sugar

Blood sugar test

Science.gov (United States)

... sugar; Blood sugar level; Fasting blood sugar; Glucose test; Diabetic screening - blood sugar test; Diabetes - blood sugar test ... The test may be done in the following ways: After you have not eaten anything for at least 8 ...

Hydrocolloids Decrease the Digestibility of Corn Starch, Soy Protein, and Skim Milk and the Antioxidant Capacity of Grape Juice.

Science.gov (United States)

Yi, Yue; Jeon, Hyeong-Ju; Yoon, Sun; Lee, Seung-Min

2015-12-01

Hydrocolloids have many applications in foods including their use in dysphagia diets. We aimed to evaluate whether hydrocolloids in foods affect the digestibility of starch and protein, and their effects on antioxidant capacity. The thickening hydrocolloids: locust bean gum and carboxymethyl cellulose, and the gel-forming agents: agar agar, konjac-glucomannan, and Hot & Soft Plus were blended with corn starch and soy protein, skim milk, or grape juice and were examined for their in vitro-digestability by comparing the reducing sugar and trichloroacetic acid (TCA)-soluble peptide, for antioxidant capacity by total polyphenol contents and the 2,2-diphenyl-1-picrylhydrazyl radical scavenging activity. The hydrocolloids resulted in a decrease in starch digestibility with the gel-forming agents. Hydrocolloids diminished TCA-soluble peptides in skim milk compared to soy protein with the exception of locust bean gum and decreased free radical scavenging capacities and total phenolic contents in grape juice. Our findings may provide evidence for the use of hydro-colloids for people at risk of nutritional deficiencies such as dysphagia patients.

Kinetic modelling of the Maillard reaction between proteins and sugars

NARCIS (Netherlands)

Brands, C.M.J.

2002-01-01

Keywords: Maillard reaction, sugar isomerisation, kinetics, multiresponse modelling, brown colour formation, lysine damage, mutagenicity, casein, monosaccharides, disaccharides, aldoses, ketoses

The aim of this thesis was to determine the kinetics of the Maillard reaction between

Biochemical and histological characterization of Agave fourcroydes Lem. plants in vitro acclimatized

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Yunel Pérez Hernández

2016-10-01

Full Text Available Henequen (Agave fourcroydes Lem. is a representative crop of the province of Matanzas, Cuba, which has been exploited extensively for the quality of its fibers. The present work aimed to characterize from the biochemical and histological point of view henequen plants cv. 'Sac Ki' in vitro acclimatized. The biochemical indicators analyzed were concentration of soluble phenols, total soluble carbohydrates, reducing sugars, soluble proteins and total chlorophylls, as well as the relative contents of flavonoids, terpins and anthocyanins. In addition, a histological analysis of the epidermis was performed and the results were compared with plants of one month in rooting culture medium. In vitro acclimatized plants were subsequently transferred to the ex vitro conditions. The contents of total soluble carbohydrates, reducing sugars and chlorophylls decreased, whereas the levels of soluble phenols, proteins and secondary metabolites were higher in relation to in vitro plants rooted for 30 days. The epidermis of the in vitro acclimatized plants showed a greater development of the stomatal apparatus that could positively affect the ex vitro acclimatization, where a survival rate of 87% and a reduction of the acclimatization time were obtained.   Keywords: chlorophyll, henequen, phenol, reducing sugar

Effect of gamma irradiation on the the quality of ginger

International Nuclear Information System (INIS)

Parvez, K.; Khawaja, M.A.; Salahuddin' Kausar, T.

1998-01-01

Ginger (Zingebar officinale) was irradiated with gamma rays (10 k and 100 k rad) and stored at room temperature in sand, in refrigerator and deep freezer. 10 k rad. dose was most appropriate because it kept the texture of ginger intact for a longer storage period without affecting its important food ingredients i.e. moisture, soluble, insoluble ash and proteins, total sugar, soluble and reducing sugar contents. (author)

Utilization of residue from cassava starch processing for production of fermentable sugar by enzymatic hydrolysis

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Luciana Reis Fontinelle SOUTO

Full Text Available Abstract The aim of this study was to characterize and perform enzymatic hydrolysis of cassava peeling residue (peel and inner peel, mainly composed of peels and small pieces. Residue was sanitized, dried at 55 °C for 24 hours and ground. The obtained flour showed pH of 4.85; 72.53 g 100 g–1 moisture; 5.18 mL 1M NaOH 100 g–1 acidity; 60.68 g 100 g–1 starch; 1.08 g 100 g–1 reducing sugar; 1.63 g 100g–1 ash; 0.86 g 100 g–1 lipid and 3.97 g 100 g–1 protein. Enzymatic hydrolysis was carried out by means of rotational central composite design, analyzing the effects of concentrations of α-amylase enzyme (10 to 50 U g starch–1, and the amyloglucosidase enzyme (80 to 400 U g starch–1 on variable responses: percent conversion of starch into reducing sugars (RSC and soluble solid content (SS. Highest values of RSC (110% and SS (12 °Brix were observed when using the maximum concentration of amyloglucosidase and throughout the concentration range of α-amylase. Enzymatic hydrolysis of cassava peel is feasible and allows the use of hydrolysate in fermentation processes for the production of various products, such as alcoholic drinks, vinegar, among others.

Utilization of residue from cassava starch processing for production of fermentable sugar by enzymatic hydrolysis

Directory of Open Access Journals (Sweden)

Luciana Reis Fontinelle SOUTO

2016-01-01

Full Text Available Abstract The aim of this study was to characterize and perform enzymatic hydrolysis of cassava peeling residue (peel and inner peel, mainly composed of peels and small pieces. Residue was sanitized, dried at 55 °C for 24 hours and ground. The obtained flour showed pH of 4.85; 72.53 g 100 g–1 moisture; 5.18 mL 1M NaOH 100 g–1 acidity; 60.68 g 100 g–1 starch; 1.08 g 100 g–1 reducing sugar; 1.63 g 100g–1 ash; 0.86 g 100 g–1 lipid and 3.97 g 100 g–1 protein. Enzymatic hydrolysis was carried out by means of rotational central composite design, analyzing the effects of concentrations of α-amylase enzyme (10 to 50 U g starch–1, and the amyloglucosidase enzyme (80 to 400 U g starch–1 on variable responses: percent conversion of starch into reducing sugars (RSC and soluble solid content (SS. Highest values of RSC (110% and SS (12 °Brix were observed when using the maximum concentration of amyloglucosidase and throughout the concentration range of α-amylase. Enzymatic hydrolysis of cassava peel is feasible and allows the use of hydrolysate in fermentation processes for the production of various products, such as alcoholic drinks, vinegar, among others.

Mitochondrial Dysfunction Plus High-Sugar Diet Provokes a Metabolic Crisis That Inhibits Growth

Science.gov (United States)

Kemppainen, Esko; George, Jack; Garipler, Görkem; Tuomela, Tea; Kiviranta, Essi; Soga, Tomoyoshi; Dunn, Cory D.; Jacobs, Howard T.

2016-01-01

The Drosophila mutant tko25t exhibits a deficiency of mitochondrial protein synthesis, leading to a global insufficiency of respiration and oxidative phosphorylation. This entrains an organismal phenotype of developmental delay and sensitivity to seizures induced by mechanical stress. We found that the mutant phenotype is exacerbated in a dose-dependent fashion by high dietary sugar levels. tko25t larvae were found to exhibit severe metabolic abnormalities that were further accentuated by high-sugar diet. These include elevated pyruvate and lactate, decreased ATP and NADPH. Dietary pyruvate or lactate supplementation phenocopied the effects of high sugar. Based on tissue-specific rescue, the crucial tissue in which this metabolic crisis initiates is the gut. It is accompanied by down-regulation of the apparatus of cytosolic protein synthesis and secretion at both the RNA and post-translational levels, including a novel regulation of S6 kinase at the protein level. PMID:26812173

Mitochondrial Dysfunction Plus High-Sugar Diet Provokes a Metabolic Crisis That Inhibits Growth.

Science.gov (United States)

Kemppainen, Esko; George, Jack; Garipler, Görkem; Tuomela, Tea; Kiviranta, Essi; Soga, Tomoyoshi; Dunn, Cory D; Jacobs, Howard T

2016-01-01

The Drosophila mutant tko25t exhibits a deficiency of mitochondrial protein synthesis, leading to a global insufficiency of respiration and oxidative phosphorylation. This entrains an organismal phenotype of developmental delay and sensitivity to seizures induced by mechanical stress. We found that the mutant phenotype is exacerbated in a dose-dependent fashion by high dietary sugar levels. tko25t larvae were found to exhibit severe metabolic abnormalities that were further accentuated by high-sugar diet. These include elevated pyruvate and lactate, decreased ATP and NADPH. Dietary pyruvate or lactate supplementation phenocopied the effects of high sugar. Based on tissue-specific rescue, the crucial tissue in which this metabolic crisis initiates is the gut. It is accompanied by down-regulation of the apparatus of cytosolic protein synthesis and secretion at both the RNA and post-translational levels, including a novel regulation of S6 kinase at the protein level.

76 FR 62339 - Domestic Sugar Program-2011-Crop Cane Sugar and Beet Sugar Marketing Allotments and Company...

Science.gov (United States)

2011-10-07

... DEPARTMENT OF AGRICULTURE Commodity Credit Corporation Domestic Sugar Program--2011-Crop Cane Sugar and Beet Sugar Marketing Allotments and Company Allocations AGENCY: Commodity Credit Corporation... the fiscal year (FY) 2012 State sugar marketing allotments and company allocations to sugarcane and...

Effect of dietary inclusion of sugar syrup on production performance, egg quality and blood biochemical parameters in laying hens

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Ahmed S. Hussein

2018-03-01

Full Text Available The effects of dietary inclusion of sugar syrup on quality of egg, cholesterol level, production performance, serum total protein and blood biochemical parameters were evaluated in laying hens. A total of 300 commercial Lohmann LSL hens (30 weeks of age were randomly distributed into 3 dietary treatments which consisted of a normal corn diet containing corn–soy and 2 diets containing 5% and 10% sugar syrup. Each treatment was replicated 5 times (n = 20. Egg production, feed intake, body weight and egg weight of laying hens fed different diets were recorded. The experiment lasted for 20 weeks. The Haugh unit scores of hens fed diets with sugar syrup were significantly increased (P < 0.05 compared with the control treatment. The sugar syrup had no significant effect on liver enzymes, total protein, blood glucose and creatinine in all treatments. The eggs laid by hens fed sugar syrup diets had lower cholesterol level (P < 0.05 compared with those laid by hens fed the control diet. Electrophoresis analysis showed that comparable electrophoretic patterns were noticed between serum proteins of treatment groups. From the results, it can be concluded that sugar syrup diets and corn diets have similar effects on feed intake, body weight, production of eggs and blood biochemical parameters in layer hens, which suggests sugar syrup can be used as an energy source for replacing part of corn in poultry layer diets. Keywords: Cholesterol, Sugar syrup diet, Egg, Egg quality, Protein

Effect of different levels of sugar on qualitative characteristics of lassi prepared from sour dahi

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Muhammad Shah Moazzem

2018-08-01

Full Text Available An investigation was carried out to develop lassi from sour dahi using different levels of sugar (10, 15, 20 and 25% and 15% water. Lassi quality was assayed through the study of physical, chemical and microbiological parameters. Results revealed that significant difference existed in overall physical score of lassi samples and the highest score was found in 15% sugar lassi whereas, the lowest score was found in 25% sugar lassi. Total solids, carbohydrate, fat, protein and ash contents differed significantly among various levels of sugar added lassi. From chemical test, it appears that, 15% sugar added lassi possess the highest fat and protein values whereas, the highest total solids and carbohydrate values posses in 25% sugar added lassi. No significant difference (p>0.05 revealed in terms of pH value and acidity percentage among lassi types. Lassi made from 10% sugar was most inferior than other levels of sugar added lassi in respect of microbiological quality- total viable count (×104 cfu/mL content was 95.67±2.08 and coliform (×10 cfu/mL content was 1.00±0.00. Considering above mentioned quality aspects, it might be resolved that lassi could be prepared successfully from sour dahi with 15% sugar keeping water level constant at 15%. [Fundam Appl Agric 2018; 3(2.000: 434-439

Chloroquine inhibits accessory cell presentation of soluble natural and synthetic protein antigens

DEFF Research Database (Denmark)

Buus, S; Werdelin, O

1984-01-01

We have studied the in vitro effect of the lysosomotrophic agent, chloroquine, on the presentation of soluble protein antigens by guinea pig accessory cells. Chloroquine inhibited the capacity of antigen-pulsed accessory cells to stimulate proliferation in appropriately primed T cells. The effect...... was time- and dose-dependent. A brief treatment solely of the accessory cells with the drug compromised their ability to stimulate primed T cells in a subsequent culture provided the accessory cells were treated with chloroquine before their exposure to the antigen. These results suggest that chloroquine...... acts on an early event in the antigen handling by accessory cells. Chloroquine is a well known inhibitor of lysosomal proteolysis, and it is likely that its effect on antigen presentation is caused by an inhibition of antigen degradation....

N-succinyl-chitosan as a drug carrier: water-insoluble and water-soluble conjugates.

Science.gov (United States)

Kato, Yoshinori; Onishi, Hiraku; Machida, Yoshiharu

2004-02-01

N-succinyl-chitosan (Suc-Chi) has favourable properties as a drug carrier such as biocompatibility, low toxicity and long-term retention in the body. It was long retained in the systemic circulation after intravenous administration, and the plasma half-lives of Suc-Chi (MW: 3.4 x 10(5); succinylation degree: 0.81 mol/sugar unit; deacetylation degree: 1.0 mol/sugar unit) were ca. 100.3h in normal mice and 43 h in Sarcoma 180-bearing mice. The biodistribution of Suc-Chi into other tissues was trace apart from the prostate and lymph nodes. The maximum tolerable dose for the intraperitoneal injection of Suc-Chi to mice was greater than 2 g/kg. The water-insoluble and water-soluble conjugates could be prepared using a water-soluble carbodiimide and mitomycin C (MMC) or using an activated ester of glutaric MMC. In vitro release characteristics of these conjugates showed similar patterns, i.e. a pH-dependent manner, except that water-insoluble conjugates showed a slightly slower release of MMC than water-soluble ones. The conjugates of MMC with Suc-Chi showed good antitumour activities against various tumours such as murine leukaemias (L1210 and P388), B16 melanoma, Sarcoma 180 solid tumour, a murine liver metastatic tumour (M5076) and a murine hepatic cell carcinoma (MH134). This review summarizes the utilization of Suc-Chi as a drug carrier for macromolecular conjugates of MMC and the therapeutic efficacy of the conjugates against various tumours.

Mitochondrial Dysfunction Plus High-Sugar Diet Provokes a Metabolic Crisis That Inhibits Growth.

Directory of Open Access Journals (Sweden)

Esko Kemppainen

Full Text Available The Drosophila mutant tko25t exhibits a deficiency of mitochondrial protein synthesis, leading to a global insufficiency of respiration and oxidative phosphorylation. This entrains an organismal phenotype of developmental delay and sensitivity to seizures induced by mechanical stress. We found that the mutant phenotype is exacerbated in a dose-dependent fashion by high dietary sugar levels. tko25t larvae were found to exhibit severe metabolic abnormalities that were further accentuated by high-sugar diet. These include elevated pyruvate and lactate, decreased ATP and NADPH. Dietary pyruvate or lactate supplementation phenocopied the effects of high sugar. Based on tissue-specific rescue, the crucial tissue in which this metabolic crisis initiates is the gut. It is accompanied by down-regulation of the apparatus of cytosolic protein synthesis and secretion at both the RNA and post-translational levels, including a novel regulation of S6 kinase at the protein level.

Enzymatic synthesis of hydrophilic undecylenic acid sugar esters and their biodegradability.

Science.gov (United States)

Raku, Takao; Kitagawa, Masaru; Shimakawa, Hiromi; Tokiwa, Yutaka

2003-01-01

To enhance water solubility of 10-undecylenic acid, which has anti-fungus, anti-bacterial and anti-virus activity, D-glucose, trehalose and sucrose were regioselectively esterified with vinyl 10-undecylenic acid ester in dimethyl formamide by a commercial protease, Bioprase conc., from Bacillus subtilis. 6-O-(10-Undecylenoyl) D-glucose, 6-O-(10-undecylenoyl) trehalose and 1'-O-(10-undecylenoyl) sucrose were obtained. The influence of structural variation by changing the sugar moiety was analyzed the surface tension and biodegradability.

Degradation of some representative polycyclic aromatic hydrocarbons by the water-soluble protein extracts from Zea mays L. cv PR32-B10.

Science.gov (United States)

Barone, Roberto; de Biasi, Margherita-Gabriella; Piccialli, Vincenzo; de Napoli, Lorenzo; Oliviero, Giorgia; Borbone, Nicola; Piccialli, Gennaro

2016-10-01

The ability of the water-soluble protein extracts from Zea mais L. cv. PR32-B10 to degrade some representative polycyclic aromatic hydrocarbons (PAHs), has been evaluated. Surface sterilized seeds of corn (Zea mais L. Pioneer cv. PR32-B10) were hydroponically cultivated in a growth chamber under no-stressful conditions. The water-soluble protein extracts isolated from maize tissues showed peroxidase, polyphenol oxidase and catalase activities. Incubation of the extracts with naphthalene, fluorene, phenanthrene and pyrene, led to formation of oxidized and/or degradation products. GC-MS and TLC monitoring of the processes showed that naphthalene, phenanthrene, fluorene and pyrene underwent 100%, 78%, 92% and 65% oxidative degradation, respectively, after 120 min. The chemical structure of the degradation products were determined by (1)H NMR and ESI-MS spectrometry. Copyright © 2016 Elsevier Ltd. All rights reserved.

Nitrogen balance and milk composition of dairy cows fed urea and soybean meal and two protein levels using sugar cane based diets

OpenAIRE

Luís Henrique Andreucci Conti; Elmeson Ferreira de Jesus; Angélica Simone Cravo Pereira; Marcos André Arcari; Kleber da Cunha Peixoto Junior; Francisco Palma Rennó; Marcos Veiga dos Santos

2014-01-01

The aim of the study was to evaluate the effect of feeding two levels of crude protein (CP) (low: 142 g CP/kg DM; and high: 156 g CP/kg DM) and two nitrogen sources (soybean meal and urea) to dairy cows using sugar cane as forage on microbial protein synthesis, the composition of the milk nitrogen fraction, nitrogen (N) balance and blood parameters. Twelve Holstein cows with an average milk yield of 22.0 ± 2.3 kg/day, and with 235 ± 40 days in milk were included in this study. The animals wer...

Changes in soluble carbohydrates during storage of Caesalpinia echinata LAM. (Brazilwood seeds, an endangered leguminous tree from the Brazilian Atlantic Forest

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I. S. Garcia

Full Text Available Caesalpinia echinata seeds stored in laboratory environmental conditions lose their viability in one month whilst under low temperatures germination is maintained for 18 months of storage. These seeds are tolerant to desiccation, keeping their viability up to 0.08 gH2O.gDW-1. Since soluble carbohydrates are believed to be involved with desiccation tolerance and seed storability, the aim of this work is to analyze the content and composition of soluble carbohydrates in C. echinata seeds during storage in paper bags (PB and glass flasks (GF at laboratory room (RT and cool (CT temperatures. In freshly harvested seeds, total soluble carbohydrates comprised approximately 10% of the dry weight, decreasing to ca. 8% over 18 months of storage at RT. In seeds stored at CT, sugars varied differently decreasing initially and being restored at the end of the analysis period. The main neutral sugars in seeds from all treatments were sucrose, fructose and glucose. Raffinose and stachyose were present as traces. Free myo-inositol and other cyclitols were also detected. The main tendency observed was the variation in levels of both glucose and fructose in relation to sucrose, the highest levels of monosaccharides which were found in seeds stored at CT. The values of glucose and fructose were practically constant in seeds stored in paper bags for 18 months at CT, decreasing consistently in the other treatments, mainly at RT. Sucrose contents remained relatively stable. Changes in soluble sugars during storage suggest that the loss of germinability of seeds of C. echinata could be associated with low levels of glucose and fructose in relation to sucrose.

Thermodynamic compatibility and interactions between Speckled Sugar bean protein and xanthan gum for production of multilayer O/W emulsion.

Science.gov (United States)

Rahmati, Nazanin Fatemeh; Koocheki, Arash; Varidi, Mehdi; Kadkhodaee, Rassoul

2018-03-01

Thermodynamic compatibility and probable interactions between Speckled Sugar been protein (SSBP) and xanthan gum for production of multilayer O/W emulsion (30% oil) were investigated. Different interactions were observed between SSBP and xanthan at different pH (3-7) including electrostatic interactions and hydrogen bonding. These interactions were predominant at pH 3. When low xanthan gum concentration (0.1%) was used, phase separation and complex coacervation observed at this pH (negative effect of interactions). However, at pH 5, only 0.1% xanthan was enough to drastically reduce non-dissolved protein and its precipitation which normally occurs at this pH. In addition, incompatibility or segregative phase behavior which normally occurs when protein and polysaccharide have same charges was not observed (positive effects of interactions). Protein-gum interactions influenced emulsion properties (zeta potential, particle size, PDI, rheology, emulsion capacity, heat stability and creaming rate). Interactions had considerable influence on emulsion shelf life and produced completely stable emulsions at all pH values. Results confirmed that SSBP-xanthan gum mixture has a high potential for production of multilayer emulsions.

Unique Features of Halophilic Proteins.

Science.gov (United States)

Arakawa, Tsutomu; Yamaguchi, Rui; Tokunaga, Hiroko; Tokunaga, Masao

2017-01-01

Proteins from moderate and extreme halophiles have unique characteristics. They are highly acidic and hydrophilic, similar to intrinsically disordered proteins. These characteristics make the halophilic proteins soluble in water and fold reversibly. In addition to reversible folding, the rate of refolding of halophilic proteins from denatured structure is generally slow, often taking several days, for example, for extremely halophilic proteins. This slow folding rate makes the halophilic proteins a novel model system for folding mechanism analysis. High solubility and reversible folding also make the halophilic proteins excellent fusion partners for soluble expression of recombinant proteins.

Impacts of Sugar Import Policy on Sugar Production in Indonesia

OpenAIRE

Suryantoro, Agustinus; Susilo, Albertus Magnus; Supriyono, Supriyono

2013-01-01

Production of sugar unful lled consumption of Indonesia society. The lack of consumption and productionhave ful lled by import. Assumption national consumption 2,7 million ton, Indonesia will import sugar in 2013predicted about 300.000 ton (Tempo.co, August, 21, 2012).The aims in general of this research are to understand the impact of sugar import policy on sugar production.Especially (1) to understand the factors that in uence sugar import price, (2) to understand impact of sugarimport pric...

A comprehensive review of the lipid cubic phase or in meso method for crystallizing membrane and soluble proteins and complexes

International Nuclear Information System (INIS)

Caffrey, Martin

2015-01-01

A comprehensive and up-to-date review of the lipid cubic phase or in meso method for crystallizing membrane and soluble proteins and complexes is reported. Recent applications of the method for in situ serial crystallography at X-ray free-electron lasers and synchrotrons are described. The lipid cubic phase or in meso method is a robust approach for crystallizing membrane proteins for structure determination. The uptake of the method is such that it is experiencing what can only be described as explosive growth. This timely, comprehensive and up-to-date review introduces the reader to the practice of in meso crystallogenesis, to the associated challenges and to their solutions. A model of how crystallization comes about mechanistically is presented for a more rational approach to crystallization. The possible involvement of the lamellar and inverted hexagonal phases in crystallogenesis and the application of the method to water-soluble, monotopic and lipid-anchored proteins are addressed. How to set up trials manually and automatically with a robot is introduced with reference to open-access online videos that provide a practical guide to all aspects of the method. These range from protein reconstitution to crystal harvesting from the hosting mesophase, which is noted for its viscosity and stickiness. The sponge phase, as an alternative medium in which to perform crystallization, is described. The compatibility of the method with additive lipids, detergents, precipitant-screen components and materials carried along with the protein such as denaturants and reducing agents is considered. The powerful host and additive lipid-screening strategies are described along with how samples that have low protein concentration and cell-free expressed protein can be used. Assaying the protein reconstituted in the bilayer of the cubic phase for function is an important element of quality control and is detailed. Host lipid design for crystallization at low temperatures and for

A comprehensive review of the lipid cubic phase or in meso method for crystallizing membrane and soluble proteins and complexes

Energy Technology Data Exchange (ETDEWEB)

Caffrey, Martin, E-mail: martin.caffrey@tcd.ie [Trinity College Dublin, Dublin (Ireland)

2015-01-01

A comprehensive and up-to-date review of the lipid cubic phase or in meso method for crystallizing membrane and soluble proteins and complexes is reported. Recent applications of the method for in situ serial crystallography at X-ray free-electron lasers and synchrotrons are described. The lipid cubic phase or in meso method is a robust approach for crystallizing membrane proteins for structure determination. The uptake of the method is such that it is experiencing what can only be described as explosive growth. This timely, comprehensive and up-to-date review introduces the reader to the practice of in meso crystallogenesis, to the associated challenges and to their solutions. A model of how crystallization comes about mechanistically is presented for a more rational approach to crystallization. The possible involvement of the lamellar and inverted hexagonal phases in crystallogenesis and the application of the method to water-soluble, monotopic and lipid-anchored proteins are addressed. How to set up trials manually and automatically with a robot is introduced with reference to open-access online videos that provide a practical guide to all aspects of the method. These range from protein reconstitution to crystal harvesting from the hosting mesophase, which is noted for its viscosity and stickiness. The sponge phase, as an alternative medium in which to perform crystallization, is described. The compatibility of the method with additive lipids, detergents, precipitant-screen components and materials carried along with the protein such as denaturants and reducing agents is considered. The powerful host and additive lipid-screening strategies are described along with how samples that have low protein concentration and cell-free expressed protein can be used. Assaying the protein reconstituted in the bilayer of the cubic phase for function is an important element of quality control and is detailed. Host lipid design for crystallization at low temperatures and for

The Malus domestica sugar transporter gene family: identifications based on genome and expression profiling related to the accumulation of fruit sugars.

Science.gov (United States)

Wei, Xiaoyu; Liu, Fengli; Chen, Cheng; Ma, Fengwang; Li, Mingjun

2014-01-01

In plants, sugar transporters are involved not only in long-distance transport, but also in sugar accumulations in sink cells. To identify members of sugar transporter gene families and to analyze their function in fruit sugar accumulation, we conducted a phylogenetic analysis of the Malus domestica genome. Expression profiling was performed with shoot tips, mature leaves, and developed fruit of "Gala" apple. Genes for sugar alcohol [including 17 sorbitol transporters (SOTs)], sucrose, and monosaccharide transporters, plus SWEET genes, were selected as candidates in 31, 9, 50, and 27 loci, respectively, of the genome. The monosaccharide transporter family appears to include five subfamilies (30 MdHTs, 8 MdEDR6s, 5 MdTMTs, 3 MdvGTs, and 4 MdpGLTs). Phylogenetic analysis of the protein sequences indicated that orthologs exist among Malus, Vitis, and Arabidopsis. Investigations of transcripts revealed that 68 candidate transporters are expressed in apple, albeit to different extents. Here, we discuss their possible roles based on the relationship between their levels of expression and sugar concentrations. The high accumulation of fructose in apple fruit is possibly linked to the coordination and cooperation between MdTMT1/2 and MdEDR6. By contrast, these fruits show low MdSWEET4.1 expression and a high flux of fructose produced from sorbitol. Our study provides an exhaustive survey of sugar transporter genes and demonstrates that sugar transporter gene families in M. domestica are comparable to those in other species. Expression profiling of these transporters will likely contribute to improving our understanding of their physiological functions in fruit formation and the development of sweetness properties.

The Malus domestica sugar transporter gene family: identifications based on genome and expression profiling related to the accumulation of fruit sugars

Directory of Open Access Journals (Sweden)

Xiaoyu eWei

2014-11-01

Full Text Available In plants, sugar transporters are involved not only in long-distance transport, but also in sugar accumulations in sink cells. To identify members of sugar transporter gene families and to analyze their function in fruit sugar accumulation, we conducted a phylogenetic analysis of the Malus domestica genome. Expression profiling was performed with shoot tips, mature leaves, and developed fruit of ‘Gala’ apple. Genes for sugar alcohol (including 17 sorbitol transporters, sucrose, and monosaccharide transporters, plus SWEET genes, were selected as candidates in 31, 9, 50, and 27 loci, respectively, of the genome. The monosaccharide transporter family appears to include five subfamilies (30 MdHTs, 8 MdEDR6s, 5 MdTMTs, 3 MdvGTs, and 4 MdpGLTs. Phylogenetic analysis of the protein sequences indicated that orthologs exist among Malus, Vitis, and Arabidopsis. Investigations of transcripts revealed that 68 candidate transporters are expressed in apple, albeit to different extents. Here, we discuss their possible roles based on the relationship between their levels of expression and sugar concentrations. The high accumulation of fructose in apple fruit is possibly linked to the coordination and cooperation between MdTMT1/2 and MdEDR6. By contrast, these fruits show low MdSWEET4.1 expression and a high flux of fructose produced from sorbitol. Our study provides an exhaustive survey of sugar transporter genes and demonstrates that sugar transporter gene families in M. domestica are comparable to those in other species. Expression profiling of these transporters will likely contribute to improving our understanding of their physiological functions in fruit formation and the development of sweetness properties.

ISOLATION AND CHARACTERIZATION OF SOLUBLE POLYSACCHARIDES FROM CALAMAGROSTIS ANGUSTIFOLIA KOM

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Xue-Fei Cao

2011-06-01

Full Text Available Sequential treatments of dewaxed Calamagrostis angustifolia Kom with water (60 ºC and 90 ºC, 70% ethanol, and 70% ethanol containing 0.2%, 1.0%, 2.0%, 4.0%, and 8.0% NaOH at a solid to liquid ratio of 1:25 (g/mL at 80 ºC for 3 h yielded 36.2% soluble polysaccharides of the dry dewaxed material. The eight polysaccharide fractions obtained were comparatively studied by sugar analysis, GPC, FT-IR, 1H and 13C-NMR, and 2D-NMR (HSQC spectroscopy. The results showed that the water-soluble polysaccharides might contain noticeable amounts of β-D-glucan, as well as some pectic substances and galactoarabinoxylan. 70% ethanol-soluble polysaccharide was mainly arabinogalactan. The five alkali-soluble hemicelluloses were mainly galactoarabinoxylans. The Ara/Xyl and Ara/Gal values of H5-H8 fractions decreased with the increment of NaOH concentration from 1.0% to 8.0%. Meanwhile, the molecular weights had a declining trend from ~60,000 to ~40,000 g/mol. The smaller sized and more branched polysaccharides tended to be extracted in the early stages under milder conditions, and the larger molecular sized and more linear hemicelluloses tended to be isolated under more highly alkaline conditions.

African Journal of Biotechnology - Vol 11, No 5 (2012)

African Journals Online (AJOL)

Enhancing the productivity of soluble green fluorescent protein through ... Effects of spermidine, proline and carbohydrate sources on somatic embryogenesis .... The dynamics of fat, protein and sugar metabolism during walnut (Juglans regia ...

Shifting Sugars and Shifting Paradigms

Science.gov (United States)

Siegal, Mark L.

2015-01-01

No organism lives in a constant environment. Based on classical studies in molecular biology, many have viewed microbes as following strict rules for shifting their metabolic activities when prevailing conditions change. For example, students learn that the bacterium Escherichia coli makes proteins for digesting lactose only when lactose is available and glucose, a better sugar, is not. However, recent studies, including three PLOS Biology papers examining sugar utilization in the budding yeast Saccharomyces cerevisiae, show that considerable heterogeneity in response to complex environments exists within and between populations. These results join similar recent results in other organisms that suggest that microbial populations anticipate predictable environmental changes and hedge their bets against unpredictable ones. The classical view therefore represents but one special case in a range of evolutionary adaptations to environmental changes that all organisms face. PMID:25688600

Shifting sugars and shifting paradigms.

Directory of Open Access Journals (Sweden)

Mark L Siegal

2015-02-01

Full Text Available No organism lives in a constant environment. Based on classical studies in molecular biology, many have viewed microbes as following strict rules for shifting their metabolic activities when prevailing conditions change. For example, students learn that the bacterium Escherichia coli makes proteins for digesting lactose only when lactose is available and glucose, a better sugar, is not. However, recent studies, including three PLOS Biology papers examining sugar utilization in the budding yeast Saccharomyces cerevisiae, show that considerable heterogeneity in response to complex environments exists within and between populations. These results join similar recent results in other organisms that suggest that microbial populations anticipate predictable environmental changes and hedge their bets against unpredictable ones. The classical view therefore represents but one special case in a range of evolutionary adaptations to environmental changes that all organisms face.

Colon-specific delivery of a probiotic-derived soluble protein ameliorates intestinal inflammation in mice through an EGFR-dependent mechanism

Science.gov (United States)

Yan, Fang; Cao, Hanwei; Cover, Timothy L.; Washington, M. Kay; Shi, Yan; Liu, LinShu; Chaturvedi, Rupesh; Peek, Richard M.; Wilson, Keith T.; Polk, D. Brent

2011-01-01

Probiotic bacteria can potentially have beneficial effects on the clinical course of several intestinal disorders, but our understanding of probiotic action is limited. We have identified a probiotic bacteria–derived soluble protein, p40, from Lactobacillus rhamnosus GG (LGG), which prevents cytokine-induced apoptosis in intestinal epithelial cells. In the current study, we analyzed the mechanisms by which p40 regulates cellular responses in intestinal epithelial cells and p40’s effects on experimental colitis using mouse models. We show that the recombinant p40 protein activated EGFR, leading to Akt activation. Activation of EGFR by p40 was required for inhibition of cytokine-induced apoptosis in intestinal epithelial cells in vitro and ex vivo. Furthermore, we developed a pectin/zein hydrogel bead system to specifically deliver p40 to the mouse colon, which activated EGFR in colon epithelial cells. Administration of p40-containing beads reduced intestinal epithelial apoptosis and disruption of barrier function in the colon epithelium in an EGFR-dependent manner, thereby preventing and treating DSS-induced intestinal injury and acute colitis. Furthermore, p40 activation of EGFR was required for ameliorating colon epithelial cell apoptosis and chronic inflammation in oxazolone-induced colitis. These data define what we believe to be a previously unrecognized mechanism of probiotic-derived soluble proteins in protecting the intestine from injury and inflammation. PMID:21606592

The ultrasound-assisted sugar extraction from sugar beet cossettes

International Nuclear Information System (INIS)

Stasiak, D.M.

2005-01-01

The aim of this work was to study the ultrasound-assisted water extraction of sugar from sugar beet cossettes. The ultrasound bath device (25 kHz, 200 W) was used. The sonication accelerated sugar diffusion at both temperatures 18 deg C and 77.6 deg C and gave the higher level of dry matter content SS (4-6 percent) and sugar content CK (7-22 percent) in juice. The SS and CK depended on time of exposition, time and temperature of extraction. In particular, the effects of 5 min ultrasound-assisted extraction were equal to 20 min extraction in traditional conditions. The shorter time, lower temperature, higher efficiency and purity of juice could be the effects of sugar extraction with ultrasound. The change of thickness of diffusion membrane, microflows in tissue as well as it's environment caused by ultrasound was the reason of acceleration of sugar extraction

Role of chlorophylls, amino acids and sugars in tea

International Nuclear Information System (INIS)

Dev Choudhury, M.N.

1980-01-01

Plucked tea shoots from clones of different varieties of tea were withered, rolled, fermented and fired by CTC and orthodox methods of manufacture. Quantitative changes in the levels of chlorophylls, amino acids and water soluble sugars during different stages of processing of tea and also changes in the contents of their degradation products were studied by feeding 14 C-labelled phenylalanine, glucose, sodium carbonate and sodium propionate to the excised shoots and subsequently analysing the products. Results are discussed and suggestions have been made about adjusting the conditions of manufacture so that the teas with desired chemical constituents are produced. (M.G.B.)

Dietary protein-to-carbohydrate ratio and added sugar as determinants of excessive gestational weight gain

DEFF Research Database (Denmark)

Maslova, Ekaterina; Halldorsson, Thorhallur I; Astrup, Arne

2015-01-01

OBJECTIVE: To examine the relation between the protein:carbohydrate (P/C) ratio and added sugar intake in pregnancy and gestational weight gain (GWG). DESIGN: A prebirth cohort including 103 119 pregnancies enrolled between 1996 and 2003. SETTING: All women in Denmark were eligible to participate...... and defined as gain in g/week. We used multivariable linear regression, including adjusting for pre-pregnancy body mass index, to calculate relative change in GWG and 95% CI. RESULTS: Average GWG was 471(224) g/week. The adjusted weight gain was 16 g/week lower (95% CI 9 to 22, p for trend ....001) in the highest (Q5) versus lowest (Q1) quintile of the P/C ratio (∼3% average reduction across the entire pregnancy). Weight gain for those with >20%E vs

Identification of the hemoglobin scavenger receptor/CD163 as a natural soluble protein in plasma

DEFF Research Database (Denmark)

Møller, Holger Jon; Peterslund, Niels Anker; Graversen, Jonas Heilskov

2002-01-01

enabled identification of a soluble plasma form of HbSR (sHbSR) having an electrophoretic mobility equal to that of recombinant HbSR consisting of the extracellular domain (scavenger receptor cysteine-rich 1-9). A sandwich enzyme-linked immunosorbent assay was established and used to measure the s...... a level of sHbSR above the range of healthy persons. Patients with myelomonocytic leukemias and pneumonia/sepsis exhibited the highest levels (up to 67.3 mg/L). In conclusion, sHbSR is an abundant plasma protein potentially valuable in monitoring patients with infections and myelomonocytic leukemia....

Structural Basis for a Ribofuranosyl Binding Protein: Insights into the Furanose Specific Transport

Energy Technology Data Exchange (ETDEWEB)

Bagaria, A.; Swaminathan, S.; Kumaran, D.; Burley, S. K.

2011-04-01

The ATP-binding cassette transporters (ABC-transporters) are members of one of the largest protein superfamilies, with representatives in all extant phyla. These integral membrane proteins utilize the energy of ATP hydrolysis to carry out certain biological processes, including translocation of various substrates across membranes and non-transport related processes such as translation of RNA and DNA repair. Typically, such transport systems in bacteria consist of an ATP binding component, a transmembrane permease, and a periplasmic receptor or binding protein. Soluble proteins found in the periplasm of gram-negative bacteria serve as the primary receptors for transport of many compounds, such as sugars, small peptides, and some ions. Ligand binding activates these periplasmic components, permitting recognition by the membrane spanning domain, which supports for transport and, in some cases, chemotaxis. Transport and chemotaxis processes appear to be independent of one another, and a few mutants of bifunctional periplasmic components reveal the absence of one or the other function. Previously published high-resolution X-ray structures of various periplasmic ligand binding proteins include Arabinose binding protein (ABP), Allose binding protein (ALBP), Glucose-galactose binding protein (GBP) and Ribose binding protein (RBP). Each of these proteins consists of two structurally similar domains connected by a three-stranded hinge region, with ligand buried between the domains. Upon ligand binding and release, various conformational changes have been observed. For RBP, open (apo) and closed (ligand bound) conformations have been reported and so for MBP. The closed/active form of the protein interacts with the integral membrane component of the system in both transport and chemotaxis. Herein, we report 1.9{angstrom} resolution X-ray structure of the R{sub f}BP periplasmic component of an ABC-type sugar transport system from Hahella chejuensis (UniProt Id Q2S7D2) bound to

Maillard-reaction-induced modification and aggregation of proteins and hardening of texture in protein bar model systems.

Science.gov (United States)

Zhou, Peng; Guo, Mufan; Liu, Dasong; Liu, Xiaoming; Labuza, Teodore P

2013-03-01

The hardening of high-protein bars causes problems in their acceptability to consumers. The objective of this study was to determine the progress of the Maillard reaction in model systems of high-protein nutritional bars containing reducing sugars, and to illustrate the influences of the Maillard reaction on the modification and aggregation of proteins and the hardening of bar matrices during storage. The progress of the Maillard reaction, glycation, and aggregation of proteins, and textural changes in bar matrices were investigated during storage at 25, 35, and 45 °C. The initial development of the Maillard reaction caused little changes in hardness; however, further storage resulted in dramatic modification of protein with formation of high-molecular-weight polymers, resulting in the hardening in texture. The replacement of reducing sugars with nonreducing ingredients such as sugar alcohols in the formula minimized the changes in texture. The hardening of high-protein bars causes problems in their acceptability to consumers. Maillard reaction is one of the mechanisms contributing to the hardening of bar matrix, particularly for the late stage of storage. The replacement of reducing sugars with nonreducing ingredients such as sugar alcohols in the formula will minimize the changes in texture. © 2013 Institute of Food Technologists®

Extremely stable soluble high molecular mass multi-protein complex with DNase activity in human placental tissue.

Directory of Open Access Journals (Sweden)

Evgeniya E Burkova

Full Text Available Human placenta is an organ which protects, feeds, and regulates the grooving of the embryo. Therefore, identification and characterization of placental components including proteins and their multi-protein complexes is an important step to understanding the placenta function. We have obtained and analyzed for the first time an extremely stable multi-protein complex (SPC, ∼ 1000 kDa from the soluble fraction of three human placentas. By gel filtration on Sepharose-4B, the SPC was well separated from other proteins of the placenta extract. Light scattering measurements and gel filtration showed that the SPC is stable in the presence of NaCl, MgCl2, acetonitrile, guanidinium chloride, and Triton in high concentrations, but dissociates efficiently in the presence of 8 M urea, 50 mM EDTA, and 0.5 M NaCl. Such a stable complex is unlikely to be a casual associate of different proteins. According to SDS-PAGE and MALDI mass spectrometry data, this complex contains many major glycosylated proteins with low and moderate molecular masses (MMs 4-14 kDa and several moderately abundant (79.3, 68.5, 52.8, and 27.2 kDa as well as minor proteins with higher MMs. The SPC treatment with dithiothreitol led to a disappearance of some protein bands and revealed proteins with lower MMs. The SPCs from three placentas efficiently hydrolyzed plasmid supercoiled DNA with comparable rates and possess at least two DNA-binding sites with different affinities for a 12-mer oligonucleotide. Progress in study of placental protein complexes can promote understanding of their biological functions.

Sap-Sugar Content of Grafted Sugar Maple Trees

Science.gov (United States)

Maurice E. Jr. Demeritt; Maurice E. Jr. Demeritt

1985-01-01

In March and April 1983, 289 and 196 young grafted sugar maple trees were tapped and evaluated for sap-sugar content. In April, sap was collected from taps both above and below the graft union. Diameter of all tapped trees at 18 inches above the ground was measured. Analysis of the data revealed that: (1) trees selected for high sugar yield cannot be reproduced by...

Effect of the combinations between pea proteins and soluble fibres on cholesterolaemia and cholesterol metabolism in rats.

Science.gov (United States)

Parolini, Cinzia; Manzini, Stefano; Busnelli, Marco; Rigamonti, Elena; Marchesi, Marta; Diani, Erika; Sirtori, Cesare R; Chiesa, Giulia

2013-10-01

Many functional foods and dietary supplements have been reported to be beneficial for the management of dyslipidaemia, one of the major risk factors for CVD. Soluble fibres and legume proteins are known to be a safe and practical approach for cholesterol reduction. The present study aimed at investigating the hypocholesterolaemic effect of the combinations of these bioactive vegetable ingredients and their possible effects on the expression of genes regulating cholesterol homeostasis. A total of six groups of twelve rats each were fed, for 28 d, Nath's hypercholesterolaemic diets, differing in protein and fibre sources, being, respectively, casein and cellulose (control), pea proteins and cellulose (pea), casein and oat fibres (oat), casein and apple pectin (pectin), pea proteins and oat fibres (pea+oat) and pea proteins and apple pectin (pea+pectin). Administration of each vegetable-containing diet was associated with lower total cholesterol concentrations compared with the control. The combinations (pea+oat and pea+pectin) were more efficacious than fibres alone in modulating cholesterolaemia ( - 53 and - 54%, respectively, at 28 d; Ppea proteins, a lower hepatic cholesterol content (Ppea proteins and oat fibres or apple pectin are extremely effective in lowering plasma cholesterol concentrations in rats and affect cellular cholesterol homeostasis by up-regulating genes involved in hepatic cholesterol turnover.

Changes in sugar content and related enzyme activities in table grape (Vitis vinifera L.) in response to foliar selenium fertilizer.

Science.gov (United States)

Zhu, Shuaimeng; Liang, Yinli; An, Xiaojuan; Kong, Fanchao; Gao, Dekai; Yin, Hongfei

2017-09-01

Spraying selenium (Se) fertilizer is an effective method for Se-enriched fruit production. Sugar content in fruit is the major factor determining berry quality. However, changes in sugar metabolism in response to Se fertilizer are unclear. Hence, this study was conducted to identify the effects of Se fertilizer on sugar metabolism and related enzyme activities of grape berries. Additionally, production of leaves with and without Se fertilizer was also investigated. Acid invertase (AI) activity, total soluble sugar and Se content in berries, and photosynthetic rate in leaves produced under Se fertilizer treatments were higher than that of control. Glucose and fructose were the primary sugars in berries, with a trace of sucrose. In both berries and leaves, neutral invertase activity was lower than AI, there was no significant difference in neutral invertase, sucrose synthase and sucrose phosphate synthase between Se fertilizer-treated and control. In berries, AI showed a significant positive correlation with glucose and fructose; also Se content was significantly correlated with sugar content. AI played an important role in the process of sugar accumulation in berries; high AI activity in berries and photosynthetic rate in leaves could explain the mechanism by which Se fertilizer affected sugar accumulation in berries. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Formation of soluble amyloid oligomers and amyloid fibrils by the multifunctional protein vitronectin

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Langen Ralf

2008-10-01

Full Text Available Abstract Background The multifunctional protein vitronectin is present within the deposits associated with Alzheimer disease (AD, age-related macular degeneration (AMD, atherosclerosis, systemic amyloidoses, and glomerulonephritis. The extent to which vitronectin contributes to amyloid formation within these plaques, which contain misfolded, amyloidogenic proteins, and the role of vitronectin in the pathophysiology of the aforementioned diseases is currently unknown. The investigation of vitronectin aggregation is significant since the formation of oligomeric and fibrillar structures are common features of amyloid proteins. Results We observed vitronectin immunoreactivity in senile plaques of AD brain, which exhibited overlap with the amyloid fibril-specific OC antibody, suggesting that vitronectin is deposited at sites of amyloid formation. Of particular interest is the growing body of evidence indicating that soluble nonfibrillar oligomers may be responsible for the development and progression of amyloid diseases. In this study we demonstrate that both plasma-purified and recombinant human vitronectin readily form spherical oligomers and typical amyloid fibrils. Vitronectin oligomers are toxic to cultured neuroblastoma and retinal pigment epithelium (RPE cells, possibly via a membrane-dependent mechanism, as they cause leakage of synthetic vesicles. Oligomer toxicity was attenuated in RPE cells by the anti-oligomer A11 antibody. Vitronectin fibrils contain a C-terminal protease-resistant fragment, which may approximate the core region of residues essential to amyloid formation. Conclusion These data reveal the propensity of vitronectin to behave as an amyloid protein and put forth the possibilities that accumulation of misfolded vitronectin may contribute to aggregate formation seen in age-related amyloid diseases.

MICROFLORA OF BEET SUGAR PRODUCTION: PROBLEMS AND SOLUTIONS

Directory of Open Access Journals (Sweden)

N. G. Kulneva

2014-01-01

Full Text Available Summary. Sugar beet is one of the strategic crops for food safety of Russia. The lack of specialized warehouse for harvest does not provide storage of roots for a long time. In the case of a thaw roots that have been defrosted unsuitable for processing. Beet and products of its processing is a good object for the development of microorganisms. Permanent microflora of sugar production are: Bacillus subtilis, Clostridium perfringes, Leuconostoc dextranicum, Torula alba, Pseudomonas fluorescens, Sarcina lutea and other kinds of microorganisms, leading to a problem processing of beet root and reduced quality of sugar. The most dangerous is the slimy bacteriosis is a bacterial disease beet caused by heterofermentative cocci of Leuconostoc (Leuconostoc mesenteroides, L. dextranicum. Product of the vital activity of microorganisms is dextran, which is synthesized from sucrose as a result of dextrany or mucous fermentation and leads to significant technological problems in processing of infected beet. Improving the efficiency of sugar production is connected with decrease in loss of quality of raw material preparation, storing and processing of sugar beet. At sugar plants use a variety of drugs that suppress the growth of pathogenic microflora, but there comes a rapid adaptation of microorganisms, therefore there is a need to implement new products to prevent damage to roots and improve the quality of produced sugar. To resolve this problem experimentally selected bactericidal drug, defined its rational concentration and conditions for the use in sugar beet production. The use of antibacterial drug in the process of extraction allows to increase the purity of diffusion juice 1.3 %, reduce the protein content in it (12.5 %; with the purity of the pure juice increases by 1.1 %, its color index is reduced by 44.7 %.

Agronomical Parameters, Sugar Profile and Antioxidant Compounds of “Catherine” Peach Cultivar Influenced by Different Plum Rootstocks

Directory of Open Access Journals (Sweden)

Carolina Font i Forcada

2014-02-01

Full Text Available The influence of seven plum rootstocks (Adesoto, Monpol, Montizo, Puebla de Soto 67 AD, PM 105 AD, St. Julien GF 655/2 and Constantí 1 on individual and total sugars, as well as on antioxidant content in fruit flesh of “Catherine” peaches, was evaluated for three years. Agronomical and basic fruit quality parameters were also determined. At twelve years after budding, significant differences were found between rootstocks for the different agronomic and fruit quality traits evaluated. The Pollizo plum rootstocks Adesoto and PM 105 AD seem to induce higher sweetness to peach fruits, based on soluble solids content, individual (sucrose, fructose and sorbitol and total sugars. A clear tendency was also observed with the rootstock Adesoto, inducing the highest content of phenolics, flavonoids, vitamin C and relative antioxidant capacity (RAC. Thus, the results of this study demonstrate the significant effect of rootstock on the sugar profile and phytochemical characteristics of peach fruits. In addition, this work shows the importance of the sugar profile, because specific sugars play an important role in peach flavour quality, as well as the studied phytochemical compounds when looking for high quality peaches with enhanced health properties.

The binding affinity of a soluble TCR-Fc fusion protein is significantly improved by crosslinkage with an anti-C{beta} antibody

Energy Technology Data Exchange (ETDEWEB)

Ozawa, Tatsuhiko; Horii, Masae; Kobayashi, Eiji [Department of Immunology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama 930-0194 (Japan); Jin, Aishun [Department of Immunology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama 930-0194 (Japan); Department of Immunology, College of Basic Medical Sciences, Harbin Medical University, 157 Baojian Road, Nangang District, Harbin 150081 (China); Kishi, Hiroyuki, E-mail: immkishi@med.u-toyama.ac.jp [Department of Immunology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama 930-0194 (Japan); Muraguchi, Atsushi [Department of Immunology, Graduate School of Medicine and Pharmaceutical Sciences, University of Toyama, 2630 Sugitani, Toyama 930-0194 (Japan)

2012-06-01

Highlights: Black-Right-Pointing-Pointer A novel soluble TCR composed of TCR V and C regions with Ig Fc region is generated. Black-Right-Pointing-Pointer TCR-Fc protein immobilized by an anti-C{beta} antibody bound to a p/MHC tetramer. Black-Right-Pointing-Pointer Binding affinity of TCR-Fc was markedly increased by binding with anti-C{beta} antibody. -- Abstract: The identification and cloning of tumor antigen-specific T cell receptors (TCRs) and the production of the soluble form of the TCR (sTCR) contributed to the development of diagnostic and therapeutic tools for cancer. Recently, several groups have reported the development of technologies for the production of sTCRs. The native sTCR has a very low binding affinity for the antigenic peptide/MHC (p/MHC) complex. In this study, we established a technology to produce high affinity, functional sTCRs. We generated a novel sTCR-Fc fusion protein composed of the TCR V and C regions of the TCR linked to the immunoglobulin (Ig) Fc region. A Western blot analysis revealed that the molecular weight of the fusion protein was approximately 60 kDa under reducing conditions and approximately 100-200 kDa under non-reducing conditions. ELISAs using various antibodies showed that the structure of each domain of the TCR-Fc protein was intact. The TCR-Fc protein immobilized by an anti-C{beta} antibody effectively bound to a p/MHC tetramer. An SPR analysis showed that the TCR-Fc protein had a low binding affinity (KD; 1.1 Multiplication-Sign 10{sup -5} M) to the p/MHC monomer. Interestingly, when the TCR-Fc protein was pre-incubated with an anti-C{beta} antibody, its binding affinity for p/MHC increased by 5-fold (2.2 Multiplication-Sign 10{sup -6} M). We demonstrated a novel method for constructing a functional soluble TCR using the Ig Fc region and showed that the binding affinity of the functional sTCR-Fc was markedly increased by an anti-C{beta} antibody, which is probably due to the stabilization of the V

Sugar beet breeding

Science.gov (United States)

Sugar beet is a recent crop developed solely for extraction of the sweetener sucrose. Breeding and improvement of Beta vulgaris for sugar has a rich historical record. Sugar beet originated from fodder beet in the 1800s, and selection has increased sugar content from 4 to 6% then to over 18% today. ...

Effect of soluble calcium and lactose on limiting flux and serum protein removal during skim milk microfiltration.

Science.gov (United States)

Adams, Michael C; Hurt, Emily E; Barbano, David M

2015-11-01

The tendency of calcium to promote microfiltration (MF) membrane fouling is well documented, but the role of lactose has not been studied. Milk protein concentrate that is 85% protein on a dry basis (MPC85) contains less calcium and lactose than skim milk. Our objectives were to determine the effects of skim milk soluble calcium and lactose concentrations on the limiting fluxes (LF) and serum protein (SP) removal factors of 0.1-µm ceramic graded permeability membranes. The MF was fed with 3 different milks: skim milk, liquid MPC85 that had been standardized to the protein content of skim milk with reverse osmosis water (MPC), and liquid MPC85 that had been standardized to the protein and lactose contents of skim milk with reverse osmosis water and lactose monohydrate (MPC+L). Retentate and permeate were continuously recycled to the feed tank. The LF for each feed was determined by increasing flux once per hour from 55 kg·m(-2)·h(-1) until flux did not increase with increasing transmembrane pressure. Temperature, pressure drop across the membrane length, and protein concentration in the retentate recirculation loop were maintained at 50°C, 220 kPa, and 8.77 ± 0.2%, respectively. Experiments were replicated 3 times and the Proc GLM procedure of SAS was used for statistical analysis. An increase in LF between skim milk (91 kg·m(-2)·h(-1)) and MPC+L (124 kg·m(-2)·h(-1)) was associated with a reduction in soluble calcium. The LF of MPC+L was lower than the LF of MPC (137 kg·m(-2)·h(-1)) due to the higher viscosity contributed by lactose. Permeates produced from the MPC and MPC+L contained more protein than the skim milk permeate due to the transfer of caseins from the micelles into the reduced-calcium sera of the MPC and MPC+L. A SP removal factor was calculated by dividing true protein in the permeate by SP in the permeate portion of the feed to describe the ease of SP passage through the membrane. No differences in SP removal factors were detected among the

The endothelial protein C receptor rs867186-GG genotype is associated with increased soluble EPCR and could mediate protection against severe malaria

DEFF Research Database (Denmark)

Shabani, Estela; Opoka, Robert O; Bangirana, Paul

2016-01-01

The endothelial protein C receptor (EPCR) appears to play an important role in Plasmodium falciparum endothelial cell binding in severe malaria (SM). Despite consistent findings of elevated soluble EPCR (sEPCR) in other infectious diseases, field studies to date have provided conflicting data abo...

Improving the process of progressive preliming in sugar production

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V. A. Golybin

2016-01-01

Full Text Available Tasks progressive preliming are to conduct neutralization, coagulation, deposition of non-sugars, the formation of a precipitate structure. This is the first operation of physical and chemical cleaning of the raw juice in the case of it in optimum conditions enables the coagulation and transferred to precipitate a substantial portion of high-molecular non-sugars, poorly soluble calcium salts that provides the cleaning effect of 14–18%, that is, up to half of the total effect on the whole station lime-carbon dioxide purification. The results preliming felt not only on the properties of the filtration of the carbonated juices, but a lso on the quality of performance of all intermediates and produced of sugar, in particular the color and turbidity of an aqueous solution, the content of the ash. In our investigations using the raw juice from sugar beet low quality found that when a uniform progressi ve change in pH of the juice during warm preliming best results for deposition rate and volume solids of the resulting precipitate are achieved when the length of 7 to 10 minutes. If it increases to 15 minutes and further reduced the deposition rate of the solid phase with a significant increase in turbidity decantate. It found that as a result of excess visit a large part of the raw juice in a progressive preliming at 60 °C increase in color prelimed juice was 30–55%, which is in agreement with the calculated values decay reducing agents in alkaline solution and formation of any additional colorants. The results confirming the feasibility of the use preddefekatsionnoy for processing the raw juice of condensed carbonate refunds.

Blockade of CD26-mediated T cell costimulation with soluble caveolin-1-Ig fusion protein induces anergy in CD4{sup +}T cells

Energy Technology Data Exchange (ETDEWEB)

Ohnuma, Kei [Department of Rheumatology and Allergy, Research Hospital, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Division of Clinical Immunology, The Advanced Clinical Research Center, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Uchiyama, Masahiko [Department of Rheumatology and Allergy, Research Hospital, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Department of Computational Intelligence and System Science, Tokyo Institute of Technology, 4259 Nagatsuta-cho, Midori-ku, Yokohama, Kanagawa 226-8503 (Japan); Hatano, Ryo; Takasawa, Wataru; Endo, Yuko [Department of Rheumatology and Allergy, Research Hospital, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Dang, Nam H. [Department of Hematologic Malignancies, Nevada Cancer Institute, Las Vegas, NV 89135 (United States); Morimoto, Chikao, E-mail: morimoto@ims.u-tokyo.ac.jp [Department of Rheumatology and Allergy, Research Hospital, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan); Division of Clinical Immunology, The Advanced Clinical Research Center, The Institute of Medical Science, The University of Tokyo, 4-6-1, Shirokanedai, Minato-ku, Tokyo 108-8639 (Japan)

2009-08-21

CD26 binds to caveolin-1 in antigen-presenting cells (APC), and that ligation of CD26 by caveolin-1 induces T cell proliferation in a TCR/CD3-dependent manner. We report herein the effects of CD26-caveolin-1 costimulatory blockade by fusion protein caveolin-1-Ig (Cav-Ig). Soluble Cav-Ig inhibits T cell proliferation and cytokine production in response to recall antigen, or allogeneic APC. Our data hence suggest that blocking of CD26-associated signaling by soluble Cav-Ig may be an effective approach as immunosuppressive therapy.

Mapping Soluble Guanylyl Cyclase and Protein Disulfide Isomerase Regions of Interaction.

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Erin J Heckler

Full Text Available Soluble guanylyl cyclase (sGC is a heterodimeric nitric oxide (NO receptor that produces cyclic GMP. This signaling mechanism is a key component in the cardiovascular system. NO binds to heme in the β subunit and stimulates the catalytic conversion of GTP to cGMP several hundred fold. Several endogenous factors have been identified that modulate sGC function in vitro and in vivo. In previous work, we determined that protein disulfide isomerase (PDI interacts with sGC in a redox-dependent manner in vitro and that PDI inhibited NO-stimulated activity in cells. To our knowledge, this was the first report of a physical interaction between sGC and a thiol-redox protein. To characterize this interaction between sGC and PDI, we first identified peptide linkages between sGC and PDI, using a lysine cross-linking reagent and recently developed mass spectrometry analysis. Together with Flag-immunoprecipitation using sGC domain deletions, wild-type (WT and mutated PDI, regions of sGC involved in this interaction were identified. The observed data were further explored with computational modeling to gain insight into the interaction mechanism between sGC and oxidized PDI. Our results indicate that PDI interacts preferentially with the catalytic domain of sGC, thus providing a mechanism for PDI inhibition of sGC. A model in which PDI interacts with either the α or the β catalytic domain is proposed.

Soluble and Membrane-Bound β-Glucosidases Are Involved in Trimming the Xyloglucan Backbone.

Science.gov (United States)

Sampedro, Javier; Valdivia, Elene R; Fraga, Patricia; Iglesias, Natalia; Revilla, Gloria; Zarra, Ignacio

2017-02-01

In many flowering plants, xyloglucan is a major component of primary cell walls, where it plays an important role in growth regulation. Xyloglucan can be degraded by a suite of exoglycosidases that remove specific sugars. In this work, we show that the xyloglucan backbone, formed by (1→4)-linked β-d-glucopyranosyl residues, can be attacked by two different Arabidopsis (Arabidopsis thaliana) β-glucosidases from glycoside hydrolase family 3. While BGLC1 (At5g20950; for β-glucosidase active against xyloglucan 1) is responsible for all or most of the soluble activity, BGLC3 (At5g04885) is usually a membrane-anchored protein. Mutations in these two genes, whether on their own or combined with mutations in other exoglycosidase genes, resulted in the accumulation of partially digested xyloglucan subunits, such as GXXG, GXLG, or GXFG. While a mutation in BGLC1 had significant effects on its own, lack of BGLC3 had only minor effects. On the other hand, double bglc1 bglc3 mutants revealed a synergistic interaction that supports a role for membrane-bound BGLC3 in xyloglucan metabolism. In addition, bglc1 bglc3 was complemented by overexpression of either BGLC1 or BGLC3 In overexpression lines, BGLC3 activity was concentrated in a microsome-enriched fraction but also was present in soluble form. Finally, both genes were generally expressed in the same cell types, although, in some cases, BGLC3 was expressed at earlier stages than BGLC1 We propose that functional specialization could explain the separate localization of both enzymes, as a membrane-bound β-glucosidase could specifically digest soluble xyloglucan without affecting the wall-bound polymer. © 2017 American Society of Plant Biologists. All Rights Reserved.

Parametric and genetic analysis of Drosophila appetitive long-term memory and sugar motivation.

Science.gov (United States)

Colomb, J; Kaiser, L; Chabaud, M-A; Preat, T

2009-06-01

Distinct forms of memory can be highlighted using different training protocols. In Drosophila olfactory aversive learning, one conditioning session triggers memory formation independently of protein synthesis, while five spaced conditioning sessions lead to the formation of long-term memory (LTM), a long-lasting memory dependent on de novo protein synthesis. In contrast, one session of odour-sugar association appeared sufficient for the fly to form LTM. We designed and tuned an apparatus that facilitates repeated discriminative conditioning by alternate presentations of two odours, one being associated with sugar, as well as a new paradigm to test sugar responsiveness (SR). Our results show that both SR and short-term memory (STM) scores increase with starvation length before conditioning. The protein dependency of appetitive LTM is independent of the repetition and the spacing of training sessions, on the starvation duration and on the strength of the unconditioned stimulus. In contrast to a recent report, our test measures an abnormal SR of radish mutant flies, which might initiate their STM and LTM phenotypes. In addition, our work shows that crammer and tequila mutants, which are deficient for aversive LTM, present both an SR and an appetitive STM defect. Using the MB247-P[switch] system, we further show that tequila is required in the adult mushroom bodies for normal sugar motivation.

Sustainability Issues and Opportunities in the Sugar and Sugar-Bioproduct Industries

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Gillian Eggleston

2015-09-01

Full Text Available Like many other industries, the sugar and sugar-bioproduct industries are facing important sustainability issues and opportunities. The relatively low and fluctuating profit for sugar, surpluses of sugar, world-wide trend to produce alternative, renewable bio-based fuels and chemicals to those derived from petroleum and reduce greenhouse gases, water- and energy-intensive factories and refineries, and increased consumer demands for sustainably manufactured products are putting pressure on the industries to diversify for sustainability. Sugar crops, including sugar and energy cane (Saccharum officinarum, sugar and energy beets (Beta vulgaris, and sweet sorghum (Sorghum bicolor L. Moench, are excellent, renewable biomass feedstocks because of their availability, their being amongst the plants that give the highest yields of carbohydrates per hectare, and high sugar contents. While much research has been focused on conversion technologies for advanced biofuels and bioproducts, attention is now focused on developing sustainable supply chains of sugar feedstocks for the new, flexible biorefineries, with customers wanting maximum feedstock reliability and quality, while minimizing cost. All biomass from sugar crops are potential feedstocks. The cogeneration of bioelectricity from bagasse and leaf residues is being increasingly manufactured in more countries and, due to the high carbon content of bagasse and leaves, can also be converted into value-added products such as biochar. Sugar crops are superior feedstocks for the production of platform chemicals for the manufacture of a range of end-products, e.g., bioplastics, chemicals, and biomaterials. In several countries and regions, green sustainability criteria are now in place and have to be met to count against national biofuel targets. Processes to convert high-fiber sugar crop biomass into biofuel have been developed but there has only been limited commercialization at the large-scale.

Role of potassium and nitrogen on sugar concentration of sugar beet

African Journals Online (AJOL)

Sugar is obtained from root of sugar beet (Beta vulgaris L.) in addition to other sources. Three important economic parameters are often considered and these are root yield, sugar concentration in root juice and total sugar yield. All the three are affected by cropping period and use of fertilisers. Existing literature suggests the ...

The Functional Quality of Soluble Recombinant Polypeptides Produced in Escherichia coli Is Defined by a Wide Conformational Spectrum▿

Science.gov (United States)

Martínez-Alonso, Mónica; González-Montalbán, Nuria; García-Fruitós, Elena; Villaverde, Antonio

2008-01-01

We have observed that a soluble recombinant green fluorescent protein produced in Escherichia coli occurs in a wide conformational spectrum. This results in differently fluorescent protein fractions in which morphologically diverse soluble aggregates abound. Therefore, the functional quality of soluble versions of aggregation-prone recombinant proteins is defined statistically rather than by the prevalence of a canonical native structure. PMID:18836021

Production of a highly active, soluble form of the cytochrome P450 reductase (CPR A) from Candida tropicalis

Science.gov (United States)

Donnelly, Mark

2006-08-01

The present invention provides soluble cytochrome p450 reductase (CPR) proteins from Candida sp. having an altered N-terminal region which results in reduced hydrophobicity of the N-terminal region. Also provided are host cells comprising the subject soluble CPR proteins. In addition, the present invention provides nucleotide and corresponding amino acid sequences for soluble CPR proteins and vectors comprising the nucleotide sequences. Methods for producing a soluble CPR, for increasing production of a dicarboxylic acid, and for detecting a cytochrome P450 are also provided.

Managing your blood sugar

Science.gov (United States)

Hyperglycemia - control; Hypoglycemia - control; Diabetes - blood sugar control; Blood glucose - managing ... sugar ( hypoglycemia ) Recognize and treat high blood sugar ( hyperglycemia ) Plan healthy meals Monitor your blood sugar (glucose) ...

ESPRIT: A Method for Defining Soluble Expression Constructs in Poorly Understood Gene Sequences.

Science.gov (United States)

Mas, Philippe J; Hart, Darren J

2017-01-01

Production of soluble, purifiable domains or multi-domain fragments of proteins is a prerequisite for structural biology and other applications. When target sequences are poorly annotated, or when there are few similar sequences available for alignments, identification of domains can be problematic. A method called expression of soluble proteins by random incremental truncation (ESPRIT) addresses this problem by high-throughput automated screening of tens of thousands of enzymatically truncated gene fragments. Rare soluble constructs are identified by experimental screening, and the boundaries revealed by DNA sequencing.

Production of Microbial Transglutaminase on Media Made from Sugar Cane Molasses and Glycerol

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Manuel Vázquez

2009-01-01

Full Text Available Transglutaminase is an enzyme that catalyses an acyl transfer reaction between γ-carboxamide groups of glutaminyl residues and lysine residues in proteins. Due to this property, this enzyme is used for enhancing textural properties of protein-rich food. The transglutaminase used as food additive is obtained by microorganisms, mainly by Streptoverticillium ladakanum. On the other hand, sugar cane molasses is a viscous liquid rich in noncrystallized carbohydrates (saccharose, glucose and fructose. In this work, the feasibility of using sugar cane molasses as a carbon source for the production of microbial transglutaminase by Streptoverticillium ladakanum NRRL 3191 has been studied. Carbon sources including sugar cane molasses (60 g of total sugars per L, glycerol (60 g/L and their mixture in a ratio of 1:1 (30 g/L of each were evaluated. Time course of microbial growth, transglutaminase activity and carbon source consumption were determined every 24 h during 120 h of fermentations at three agitation speeds (200, 300 or 400 rpm. The results showed that with the increase in agitation speed, the biomass concentration increased up to 8.39 g/L in the medium containing sugar cane molasses alone or the mixture of molasses and glycerol. The highest transglutaminase activity was obtained at 400 rpm in the medium containing a mixture of molasses and glycerol, reaching 0.460 U/mL, while in the medium containing sugar cane molasses alone, the activity was 0.240 U/mL, and using glycerol alone it was 0.250 U/mL. These results show that sugar cane molasses is a suitable medium for transglutaminase production when it is combined with glycerol.

Abalone water-soluble matrix for self-healing biomineralization of tooth defects

Energy Technology Data Exchange (ETDEWEB)

Wen, Zhenliang [Institute of Biomedical and Pharmaceutical Technology, Fuzhou University, Fuzhou 350002 (China); Chen, Jingdi, E-mail: ibptcjd@fzu.edu.cn [Institute of Biomedical and Pharmaceutical Technology, Fuzhou University, Fuzhou 350002 (China); Wang, Hailiang [The Affiliated Stomatological Hospital, Fujian Medical University, Fuzhou 350002 (China); Zhong, Shengnan; Hu, Yimin; Wang, Zhili [Institute of Biomedical and Pharmaceutical Technology, Fuzhou University, Fuzhou 350002 (China); Zhang, Qiqing, E-mail: zhangqiq@126.com [Institute of Biomedical and Pharmaceutical Technology, Fuzhou University, Fuzhou 350002 (China); Institute of Biomedical Engineering, Chinese Academy of Medical Science & Peking Union Medical College, Tianjin 300192 (China)

2016-10-01

Enamel cannot heal by itself if damaged. Hydroxyapatite (HAP) is main component of human enamel. Formation of enamel-like materials for healing enamel defects remains a challenge. In this paper, we successfully isolated the abalone water-soluble matrix (AWSM) with 1.53 wt% the abalone water-soluble protein (AWSPro) and 2.04 wt% the abalone water-soluble polysaccharide (AWSPs) from abandoned abalone shell, and self-healing biomineralization of tooth defects was successfully achieved in vitro. Based on X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), hot field emission scanning electron microscopy (HFESEM) and energy dispersive spectrometer (EDS) analysis, the results showed that the AWSM can efficiently induce remineralization of HAP. The enamel-like HAP was successfully achieved onto etched enamel's surface due to the presence of the AWSM. Moreover, the remineralized effect of eroded enamel was growing with the increase of the AWSM. This study provides a solution to the resource waste and environmental pollution caused by abandoned abalone shell, and we provides a new method for self-healing remineralization of enamel defects by AWSM and develops a novel dental material for potential clinical dentistry application. - Graphical abstract: In this paper, we successfully isolated the abalone water-soluble matrix (AWSM) with 1.53 wt% abalone water-soluble protein (AWSPro) and 2.04 wt% abalone water-soluble polysaccharide (AWSPs) from abandoned abalone shell, and self-healing biomineralization of tooth defects was successfully achieved in vitro by self-organized. Display Omitted - Highlights: • Provides a solution to the resource waste and environmental pollution caused by abandoned abalone shell. • The abalone shell water-soluble matrix contains protein and polysaccharide. • The abalone water-soluble matrix can efficiently induce remineralization of HAP by self-organized. • Achieved self-healing biomineralization of tooth defects in

Abalone water-soluble matrix for self-healing biomineralization of tooth defects

International Nuclear Information System (INIS)

Wen, Zhenliang; Chen, Jingdi; Wang, Hailiang; Zhong, Shengnan; Hu, Yimin; Wang, Zhili; Zhang, Qiqing

2016-01-01

Enamel cannot heal by itself if damaged. Hydroxyapatite (HAP) is main component of human enamel. Formation of enamel-like materials for healing enamel defects remains a challenge. In this paper, we successfully isolated the abalone water-soluble matrix (AWSM) with 1.53 wt% the abalone water-soluble protein (AWSPro) and 2.04 wt% the abalone water-soluble polysaccharide (AWSPs) from abandoned abalone shell, and self-healing biomineralization of tooth defects was successfully achieved in vitro. Based on X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), hot field emission scanning electron microscopy (HFESEM) and energy dispersive spectrometer (EDS) analysis, the results showed that the AWSM can efficiently induce remineralization of HAP. The enamel-like HAP was successfully achieved onto etched enamel's surface due to the presence of the AWSM. Moreover, the remineralized effect of eroded enamel was growing with the increase of the AWSM. This study provides a solution to the resource waste and environmental pollution caused by abandoned abalone shell, and we provides a new method for self-healing remineralization of enamel defects by AWSM and develops a novel dental material for potential clinical dentistry application. - Graphical abstract: In this paper, we successfully isolated the abalone water-soluble matrix (AWSM) with 1.53 wt% abalone water-soluble protein (AWSPro) and 2.04 wt% abalone water-soluble polysaccharide (AWSPs) from abandoned abalone shell, and self-healing biomineralization of tooth defects was successfully achieved in vitro by self-organized. Display Omitted - Highlights: • Provides a solution to the resource waste and environmental pollution caused by abandoned abalone shell. • The abalone shell water-soluble matrix contains protein and polysaccharide. • The abalone water-soluble matrix can efficiently induce remineralization of HAP by self-organized. • Achieved self-healing biomineralization of tooth defects in vitro.

Segmental isotope labeling of proteins for NMR structural study using a protein S tag for higher expression and solubility

International Nuclear Information System (INIS)

Kobayashi, Hiroshi; Swapna, G. V. T.; Wu, Kuen-Phon; Afinogenova, Yuliya; Conover, Kenith; Mao, Binchen; Montelione, Gaetano T.; Inouye, Masayori

2012-01-01

A common obstacle to NMR studies of proteins is sample preparation. In many cases, proteins targeted for NMR studies are poorly expressed and/or expressed in insoluble forms. Here, we describe a novel approach to overcome these problems. In the protein S tag-intein (PSTI) technology, two tandem 92-residue N-terminal domains of protein S (PrS 2 ) from Myxococcus xanthus is fused at the N-terminal end of a protein to enhance its expression and solubility. Using intein technology, the isotope-labeled PrS 2 -tag is replaced with non-isotope labeled PrS 2 -tag, silencing the NMR signals from PrS 2 -tag in isotope-filtered 1 H-detected NMR experiments. This method was applied to the E. coli ribosome binding factor A (RbfA), which aggregates and precipitates in the absence of a solubilization tag unless the C-terminal 25-residue segment is deleted (RbfAΔ25). Using the PrS 2 -tag, full-length well-behaved RbfA samples could be successfully prepared for NMR studies. PrS 2 (non-labeled)-tagged RbfA (isotope-labeled) was produced with the use of the intein approach. The well-resolved TROSY-HSQC spectrum of full-length PrS 2 -tagged RbfA superimposes with the TROSY-HSQC spectrum of RbfAΔ25, indicating that PrS 2 -tag does not affect the structure of the protein to which it is fused. Using a smaller PrS-tag, consisting of a single N-terminal domain of protein S, triple resonance experiments were performed, and most of the backbone 1 H, 15 N and 13 C resonance assignments for full-length E. coli RbfA were determined. Analysis of these chemical shift data with the Chemical Shift Index and heteronuclear 1 H– 15 N NOE measurements reveal the dynamic nature of the C-terminal segment of the full-length RbfA protein, which could not be inferred using the truncated RbfAΔ25 construct. CS-Rosetta calculations also demonstrate that the core structure of full-length RbfA is similar to that of the RbfAΔ25 construct.

Efficient soluble expression of disulfide bonded proteins in the cytoplasm of Escherichia coli in fed-batch fermentations on chemically defined minimal media.

Science.gov (United States)

Gąciarz, Anna; Khatri, Narendar Kumar; Velez-Suberbie, M Lourdes; Saaranen, Mirva J; Uchida, Yuko; Keshavarz-Moore, Eli; Ruddock, Lloyd W

2017-06-15

The production of recombinant proteins containing disulfide bonds in Escherichia coli is challenging. In most cases the protein of interest needs to be either targeted to the oxidizing periplasm or expressed in the cytoplasm in the form of inclusion bodies, then solubilized and re-folded in vitro. Both of these approaches have limitations. Previously we showed that soluble expression of disulfide bonded proteins in the cytoplasm of E. coli is possible at shake flask scale with a system, known as CyDisCo, which is based on co-expression of a protein of interest along with a sulfhydryl oxidase and a disulfide bond isomerase. With CyDisCo it is possible to produce disulfide bonded proteins in the presence of intact reducing pathways in the cytoplasm. Here we scaled up production of four disulfide bonded proteins to stirred tank bioreactors and achieved high cell densities and protein yields in glucose fed-batch fermentations, using an E. coli strain (BW25113) with the cytoplasmic reducing pathways intact. Even without process optimization production of purified human single chain IgA 1 antibody fragment reached 139 mg/L and hen avidin 71 mg/L, while purified yields of human growth hormone 1 and interleukin 6 were around 1 g/L. Preliminary results show that human growth hormone 1 was also efficiently produced in fermentations of W3110 strain and when glucose was replaced with glycerol as the carbon source. Our results show for the first time that efficient production of high yields of soluble disulfide bonded proteins in the cytoplasm of E. coli with the reducing pathways intact is feasible to scale-up to bioreactor cultivations on chemically defined minimal media.

Dehydrated chicory pulp as an alternative soluble fibre source in diets for growing rabbits

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L. Maertens

2014-06-01

Full Text Available Soluble fibre (SF is an important nutrient to enhance fermentative activity and gut health in rabbits. The main source of SF in rabbit diets is sugar beet pulp (SBP, whereas, due to its high content of SF (34%, dried chicory pulp (ChP could be an alternative to SBP. In a fattening trial with 192 hybrid weanlings 32 d old weighing 837±45 g, chicory pulp was used in replacement of SBP to study effects on production performances and slaughter characteristics. Rabbits were fed one of 4 iso-energetic (9.65 MJ digestible energy/kg and isonitrogenous (15.6% crude protein diets: a negative control (NC diet with a low dietary SF content (7.3%, a positive control diet with quite a high SBP level (13.5% and SF content (10.6% and 2 diets with respectively 10% and 20% of chicory pulp (ChP10: 9.9% SF and ChP20: 13.7% SF. The SF content was measured as the difference between total dietary fibre and neutral detergent fibre, the latter corrected for ash and protein content. Each dietary treatment consisted of 12 replicates of 4 rabbits. Weight gain was high (on av. 54 g/d and comparable for the NC, SBP and ChP diets. However, feed conversion ratio was improved (P<0.05 with the ChP20 diet compared to the NC diet (2.88 vs. 2.97. Mortality was low and not influenced by the dietary treatment. Slaughter data were very similar and no effect of the SF level on caecal weight or slaughter yield was observed. It was concluded that chicory pulp is a good alternative soluble fibre source in balanced diets for rabbits and can be used at least up to 20% inclusion rate.

Intrinsic nitric oxide regulates the taste response of the sugar receptor cell in the blowfly, Phormia regina.

Science.gov (United States)

Murata, Yoshihiro; Mashiko, Masashi; Ozaki, Mamiko; Amakawa, Taisaku; Nakamura, Tadashi

2004-01-01

The taste organ in insects is a hair-shaped taste sensory unit having four functionally differentiated contact chemoreceptor cells. In the blowfly, Phormia regina, cGMP has been suggested to be a second messenger for the sugar receptor cell. Generally, cGMP is produced by membranous or soluble guanylyl cyclase (sGC), which can be activated by nitric oxide (NO). In the present paper, we electrophysiologically showed that an NO scavenger, 2-phenyl-4,4,5,5-tetramethylimidazoline-3-oxide-1-oxyl (PTIO), an NO donor, 1-hydroxy-2-oxo-3-(N-methyl-3-aminopropyl)-3-methyl-1-triazene (NOC 7) or an NO synthase (NOS) inhibitor, NG-nitro-L-arginine methyl ester (L-NAME) specifically affected the response in the sugar receptor cell, but not in other receptor cells. PTIO, when introduced into the receptor cells in a sensillum aided by sodium deoxycholate (DOC, pH 7.2), depressed the response of sugar receptor cells to sucrose but did not affect those of the salt or water receptor cells. NOC 7, given extracellularly, latently induced the response of sugar receptor cells; and L-NAME, when introduced into the receptor cells, depressed the response of sugar receptor cells. The results clearly suggest that NO, which may be produced by intrinsic NOS in sugar receptor cells, participates in the transduction cascade of these cells in blowfly.

Sugar-Fat Seesaw: A Systematic Review of the Evidence

Science.gov (United States)

Sadler, Michele Jeanne; McNulty, Helene; Gibson, Sigrid

2015-01-01

Further to reports of a reciprocal relationship between sugar and fat intakes, this review aimed to provide an in-depth analysis and to determine the likely influence of this relationship on the achievement of population dietary guidelines. Using systematic methods, relevant literature was selected according to preset criteria. A strong and consistent inverse association was found between total sugars and total fat intakes expressed as percentage energy. Fewer studies considered absolute intakes and these reported a positive relationship, which may be influenced by confounding with energy intakes. Evidence for an inverse relationship between percentage energy from fat and extrinsic sugars was weaker and less consistent than for fat and total sugars. Reciprocal relationships were also observed for sugar-saturated fat, sugar−protein, sugar−alcohol, and sugar−starch expressed as percentage energy. Under-reporting of dietary intakes had no major influence on the findings. This review confirms the existence of the sugar−fat seesaw on a percentage energy basis and concludes that it is most likely explained by a combination of mathematical and food compositional effects. This finding is relevant because dietary guidelines are expressed as percentage energy and implies that at the population level multiple guidelines may be difficult to achieve in practice. PMID:24915391

Expression by Streptomyces lividans of the Rat α Integrin CD11b A-Domain as a Secreted and Soluble Recombinant Protein

Directory of Open Access Journals (Sweden)

Dorra Zouari Ayadi

2007-01-01

Full Text Available We already reported the use of a long synthetic signal peptide (LSSP to secrete the Streptomyces sp. TO1 amylase by Streptomyces lividans strain. We herein report the expression and secretion of the rat CD11b A-domain using the same LSSP and S. lividans as host strain. We have used the Escherichia coli/Streptomyces shuttle vector pIJ699 for the cloning of the A-domain DNA sequence downstream of LSSP and under the control of the constitutive ermE-up promoter of Streptomyces erythraeus. Using this construct and S. lividans as a host strain, we achieved the expression of 8 mg/L of soluble secreted recombinant form of the A-domain of the rat leukocyte β2 integrin CD11/CD18 alpha M subunit (CD11b. This secreted recombinant CD11b A-domain reacted with a function blocking antibody showing that this protein is properly folded and probably functional. These data support the capability of Streptomyces to produce heterologous recombinant proteins as soluble secreted form using the “LSSP” synthetic signal peptide.

Sugars in atmospheric aerosols over the Eastern Mediterranean

Science.gov (United States)

Theodosi, Christina; Panagiotopoulos, Christos; Nouara, Amel; Zarmpas, Pavlos; Nicolaou, Panagiota; Violaki, Kalliopi; Kanakidou, Maria; Sempéré, Richard; Mihalopoulos, Nikolaos

2017-04-01

The role of biomass combustion and primary bio-particles in atmospheric PM10 aerosols in the Eastern Mediterranean over a two-year period was estimated by studying sugar tracers. Sugar concentrations ranged from 6 to 334 ng m-3, while their contributions to the organic carbon (OC) and water soluble organic carbon (WSOC) pools were 3 and 11%, respectively. Over the studied period, glucose and levoglucosan were the two most abundant sugars accounting equally about 25% of the total sugar concentration in PM10 aerosols whereas fructose, sucrose, and mannitol represented 18%, 15% and 10%, respectively. Primary saccharides (glucose, fructose, and sucrose) peaked at the beginning of spring (21, 17 and 15 ng m-3, respectively), indicating significant contributions of bioaerosols to the total organic aerosol mass. On the other hand, higher concentrations of anhydrosugars (burning biomass tracers including levoglucosan, mannosan and galactosan) were recorded in winter (19, 1.4 and 0.2 ng m-3, respectively) than in summer (9.1, 1.1 and 0.5 ng m-3, respectively). Levoglucosan was the dominant monosaccharide in winter (37% of total sugars) with less contribution in summer (19%) probably in relation with enhanced photochemical oxidation reactions by hydroxyl (ṡOH) radicals impacting anhydrosugars. We estimate that atmospheric oxidation by ṡOH decreases levoglucosan levels by 54% during summer. Biomass burning, based on levoglucosan observations, was estimated to contribute up to 13% to the annual average OC measured at Finokalia. Annual OC, WSOC, and carbohydrate dry deposition fluxes for the studied period were estimated to 414, 175, and 9 mg C m-2 y-1, respectively. Glucose and levoglucosan accounted for 34% and 2% of the total sugar fluxes. According to our estimations, atmospheric OC and WSOC inputs account for ˜0.70% of the carbon produced by annual primary production (PP) in the Cretan Sea. Considering the entire Mediterranean, dry deposition of OC could provide at

Identification, cloning and characterization of sis7 and sis10 sugar-insensitive mutants of Arabidopsis

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Biddle Kelly D

2008-10-01

Full Text Available Abstract Background The levels of soluble sugars, such as glucose and sucrose, help regulate many plant metabolic, physiological and developmental processes. Genetic screens are helping identify some of the loci involved in plant sugar response and reveal extensive cross-talk between sugar and phytohormone response pathways. Results A forward genetic screen was performed to identify mutants with increased resistance to the inhibitory effects of high levels of exogenous sugars on early Arabidopsis seedling development. The positional cloning and characterization of two of these sugar insensitive (sis mutants, both of which are also involved in abscisic acid (ABA biosynthesis or response, are reported. Plants carrying mutations in SIS7/NCED3/STO1 or SIS10/ABI3 are resistant to the inhibitory effects of high levels of exogenous Glc and Suc. Quantitative RT-PCR analyses indicate transcriptional upregulation of ABA biosynthesis genes by high concentrations of Glc in wild-type germinating seeds. Gene expression profiling revealed that a significant number of genes that are expressed at lower levels in germinating sis7-1/nced3-4/sto1-4 seeds than in wild-type seeds are implicated in auxin biosynthesis or transport, suggesting cross-talk between ABA and auxin response pathways. The degree of sugar insensitivity of different sis10/abi3 mutant seedlings shows a strong positive correlation with their level of ABA insensitivity during seed germination. Conclusion Mutations in the SIS7/NCED3/STO1 gene, which is primarily required for ABA biosynthesis under drought conditions, confer a sugar-insensitive phenotype, indicating that a constitutive role in ABA biosynthesis is not necessary to confer sugar insensitivity. Findings presented here clearly demonstrate that mutations in ABI3 can confer a sugar-insensitive phenotype and help explain previous, mixed reports on this topic by showing that ABA and sugar insensitivity exhibit a strong positive correlation in

alpha isoforms of soluble and membrane-linked folate-binding protein in human blood

DEFF Research Database (Denmark)

Hoier-Madsen, M.; Holm, J.; Hansen, S.I.

2008-01-01

supported the hypothesis that serum FBP (29 kDa) mainly originates from neutrophils. The presence of FBP/FR alpha isoforms were established for the first time in human blood using antibodies specifically directed against human milk FBP alpha. The alpha isoforms identified on erythrocyte membranes......, and in granulocytes and serum, only constituted an almost undetectable fraction of the functional FBP The FBP alpha in neutrophil granulocytes was identified as a cytoplasmic component by indirect immunofluorescence. Gel filtration of serum revealed a peak of FBP alpha (>120 kDa), which could represent receptor...... fragments from decomposed erythrocytes and granulocytes. The soluble FBPs may exert bacteriostatic effects and protect folates in plasma from biological degradation, whereas FRs on the surface of blood cells could be involved in intracellular folate uptake or serve as signal proteins. The latter receptors...

Dual effect of soluble materials in pretreated lignocellulose on simultaneous saccharification and co-fermentation process for the bioethanol production.

Science.gov (United States)

Qin, Lei; Li, Xia; Liu, Li; Zhu, Jia-Qing; Guan, Qi-Man; Zhang, Man-Tong; Li, Wen-Chao; Li, Bing-Zhi; Yuan, Ying-Jin

2017-01-01

In this study, wash liquors isolated from ethylenediamine and dry dilute acid pretreated corn stover were used to evaluate the effect of soluble materials in pretreated biomass on simultaneous saccharification and co-fermentation (SSCF) for ethanol production, respectively. Both of the wash liquors had different impacts on enzymatic hydrolysis and fermentation. Enzymatic conversions of glucan and xylan monotonically decreased as wash liquor concentration increased. Whereas, with low wash liquor concentrations, xylose consumption rate, cell viability and ethanol yield were maximally stimulated in fermentation without nutrient supplementary. Soluble lignins were found as the key composition which promoted sugars utilization and cell viability without nutrient supplementary. The dual effects of soluble materials on enzymatic hydrolysis and fermentation resulted in the reduction of ethanol yield as soluble materials increased in SSCF. Copyright © 2016 Elsevier Ltd. All rights reserved.

NUTRITIONAL CHARACTERISTICS OF EMMER WHEAT VARIETIES

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Magdaléna Lacko - Bartošová

2015-02-01

Full Text Available The objective of this study was to evaluate the nutritional compounds (fat, sugars, crude protein, soluble fiber, ash and starch of four emmer wheat varieties grown under the conditions of organic farming system. The experiment was established on Scientific Research base Dolná Malanta, near Nitra in Slovakia during 2010 – 2011 and 2011 – 2012 growing seasons. Nutritional parameters, except crude protein content, were not influenced by the variety and weather conditions. Agnone variety had the highest content of fat, crude protein and starch but the lowest content of soluble dietary fiber. The lowest values of fat, crude protein had Molise sel Colli variety; Farvento variety had the lowest sugars and starch content. Emmer wheat as ancient wheat has a unique composition in secondary components, such as starch, which may play a role as functional food ingredients.

Expression and purification of short hydrophobic elastin-like polypeptides with maltose-binding protein as a solubility tag.

Science.gov (United States)

Bataille, Laure; Dieryck, Wilfrid; Hocquellet, Agnès; Cabanne, Charlotte; Bathany, Katell; Lecommandoux, Sébastien; Garbay, Bertrand; Garanger, Elisabeth

2015-06-01

Elastin-like polypeptides (ELPs) are biodegradable polymers with interesting physico-chemical properties for biomedical and biotechnological applications. The recombinant expression of hydrophobic elastin-like polypeptides is often difficult because they possess low transition temperatures, and therefore form aggregates at sub-ambient temperatures. To circumvent this difficulty, we expressed in Escherichia coli three hydrophobic ELPs (VPGIG)n with variable lengths (n=20, 40, and 60) in fusion with the maltose-binding protein (MBP). Fusion proteins were soluble and yields of purified MBP-ELP ranged between 66 and 127mg/L culture. After digestion of the fusion proteins by enterokinase, the ELP moiety was purified by using inverse transition cycling. The purified fraction containing ELP40 was slightly contaminated by traces of undigested fusion protein. Purification of ELP60 was impaired because of co-purification of the MBP tag during inverse transition cycling. ELP20 was successfully purified to homogeneity, as assessed by gel electrophoresis and mass spectrometry analyses. The transition temperature of ELP20 was measured at 15.4°C in low salt buffer. In conclusion, this method can be used to produce hydrophobic ELP of low molecular mass. Copyright © 2015 Elsevier Inc. All rights reserved.

Polysaccharide components from the scape of Musa paradisiaca: main structural features of water-soluble polysaccharide component.

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Anjaneyalu, Y V; Jagadish, R L; Raju, T S

1997-06-01

Polysaccharide components present in the pseudo-stem (scape) of M. paradisiaca were purified from acetone powder of the scape by delignification followed by extraction with aqueous solvents into water soluble polysaccharide (WSP), EDTA-soluble polysaccharide (EDTA-SP), alkali-soluble polysaccharide (ASP) and alkali-insoluble polysaccharide (AISP) fractions. Sugar compositional analysis showed that WSP and EDTA-SP contained only D-Glc whereas ASP contained D-Glc, L-Ara and D-Xyl in approximately 1:1:10 ratio, respectively, and AISP contained D-Glc, L-Ara and D-Xyl in approximately 10:1:2 ratio, respectively. WSP was further purified by complexation with iso-amylalcohol and characterized by specific rotation, IR spectroscopy, Iodine affinity, ferricyanide number, blue value, hydrolysis with alpha-amylase and glucoamylase, and methylation linkage analysis, and shown to be a amylopectin type alpha-D-glucan.

Grapevine tissues and phenology differentially affect soluble carbohydrates determination by capillary electrophoresis.

Science.gov (United States)

Moreno, Daniela; Berli, Federico; Bottini, Rubén; Piccoli, Patricia N; Silva, María F

2017-09-01

Soluble carbohydrates distribution depends on plant physiology and, among other important factors, determines fruit yield and quality. In plant biology, the analysis of sugars is useful for many purposes, including metabolic studies. Capillary electrophoresis (CE) proved to be a powerful green separation technique with minimal sample preparation, even in complex plant tissues, that can provide high-resolution efficiency. Matrix effect refers to alterations in the analytical response caused by components of a sample other than the analyte of interest. Thus, the assessment and reduction of the matrix factor is fundamental for metabolic studies in different matrices. The present study evaluated the source and levels of matrix effects in the determination of most abundant sugars in grapevine tissues (mature and young leaves, berries and roots) at two phenological growth stages. Sucrose was the sugar that showed the least matrix effects, while fructose was the most affected analyte. Based on plant tissues, young leaves presented the smaller matrix effects, irrespectively of the phenology. These changes may be attributed to considerable differences at chemical composition of grapevine tissues with plant development. Therefore, matrix effect should be an important concern for plant metabolomics. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

A rare sugar, d-allose, confers resistance to rice bacterial blight with upregulation of defense-related genes in Oryza sativa.

Science.gov (United States)

Kano, Akihito; Gomi, Kenji; Yamasaki-Kokudo, Yumiko; Satoh, Masaru; Fukumoto, Takeshi; Ohtani, Kouhei; Tajima, Shigeyuki; Izumori, Ken; Tanaka, Keiji; Ishida, Yutaka; Tada, Yasuomi; Nishizawa, Yoko; Akimitsu, Kazuya

2010-01-01

We investigated responses of rice plant to three rare sugars, d-altrose, d-sorbose, and d-allose, due to establishment of mass production methods for these rare sugars. Root growth and shoot growth were significantly inhibited by d-allose but not by the other rare sugars. A large-scale gene expression analysis using a rice microarray revealed that d-allose treatment causes a high upregulation of many defense-related, pathogenesis-related (PR) protein genes in rice. The PR protein genes were not upregulated by other rare sugars. Furthermore, d-allose treatment of rice plants conferred limited resistance of the rice against the pathogen Xanthomonas oryzae pv. oryzae but the other tested sugars did not. These results indicate that d-allose has a growth inhibitory effect but might prove to be a candidate elicitor for reducing disease development in rice.

Thermal, mechanical, and physical properties of seaweed/sugar palm fibre reinforced thermoplastic sugar palm Starch/Agar hybrid composites.

Science.gov (United States)

Jumaidin, Ridhwan; Sapuan, Salit M; Jawaid, Mohammad; Ishak, Mohamad R; Sahari, Japar

2017-04-01

The aim of this research is to investigate the effect of sugar palm fibre (SPF) on the mechanical, thermal and physical properties of seaweed/thermoplastic sugar palm starch agar (TPSA) composites. Hybridized seaweed/SPF filler at weight ratio of 25:75, 50:50 and 75:25 were prepared using TPSA as a matrix. Mechanical, thermal and physical properties of hybrid composites were carried out. Obtained results indicated that hybrid composites display improved tensile and flexural properties accompanied with lower impact resistance. The highest tensile (17.74MPa) and flexural strength (31.24MPa) was obtained from hybrid composite with 50:50 ratio of seaweed/SPF. Good fibre-matrix bonding was evident in the scanning electron microscopy (SEM) micrograph of the hybrid composites' tensile fracture. Fourier transform infrared spectroscopy (FT-IR) analysis showed increase in intermolecular hydrogen bonding following the addition of SPF. Thermal stability of hybrid composites was enhanced, indicated by a higher onset degradation temperature (259°C) for 25:75 seaweed/SPF composites than the individual seaweed composites (253°C). Water absorption, thickness swelling, water solubility, and soil burial tests showed higher water and biodegradation resistance of the hybrid composites. Overall, the hybridization of SPF with seaweed/TPSA composites enhances the properties of the biocomposites for short-life application; that is, disposable tray, plate, etc. Copyright © 2017 Elsevier B.V. All rights reserved.

[Effects of exogenous nitric oxide on physiological characteristics of longan (Dimocarpus longana) seedlings under acid rain stress].

Science.gov (United States)

Liu, Jian-fu; Wang, Ming-yuan; Yang, Chen; Zhu, Ai-jun

2013-08-01

This paper studied the effects of exogenous nitric oxide donor sodium nitroprusside (SNP) on the chlorophyll content, antioxidant enzyme activities, and osmotic regulation substances of longan (Dimocarpus longana 'Fuyan') seedlings under acid rain (pH 3.0) stress. Under the acid rain stress, the seedling leaf superoxide dismutase (SOD), peroxidase (POD) and catalase (CAT) activities and chlorophyll, soluble protein and soluble sugar contents decreased obviously, while the leaf malondialdedyde content had a remarkable increase, suggesting the toxic effect of the acid rain on the seedlings. Exogenous nitric oxide had dual nature on the physiological characteristics of longan seedlings under acid rain stress. Applying 0.1-0.5 mmol x L(-1) of SNP improved the SOD, POD and CAT activities and the chlorophyll, soluble protein and soluble sugar contents significantly, and decreased the malondialdedyde content. Low concentrations SNP reduced the oxidative damage caused by the acid rain stress, and 0.5 mmol x L(-1) of SNP had the best effect. Under the application of 0.5 mmol x L(-1) of SNP, the total chlorophyll, soluble protein, and soluble sugar contents and the SOD, POD and CAT activities increased by 76.0%, 107.0%, 216.1%, 150. 0%, 350.9% and 97.1%, respectively, and the malondialdedyde content decreased by 46.4%. It was suggested that low concentration (0.1-0.5 mmol x L(-1)) SNP could alleviate the toxic effect of acid rain stress on longan seedlings via activating the leaf antioxidant enzyme activities and reducing oxidative stress, while high concentration SNP (1.0 mmol x L(-1)) lowered the mitigation effect.

Coarse-grain modelling of protein-protein interactions

NARCIS (Netherlands)

Baaden, Marc; Marrink, Siewert J.

2013-01-01

Here, we review recent advances towards the modelling of protein-protein interactions (PPI) at the coarse-grained (CG) level, a technique that is now widely used to understand protein affinity, aggregation and self-assembly behaviour. PPI models of soluble proteins and membrane proteins are

The Truth about Sugar.

Science.gov (United States)

Yeung, C Albert; Goodfellow, Ashley; Flanagan, Louise

2015-01-01

Sugars are used by the industry to enhance the attractiveness of foods and drinks. These added sugars, or 'free sugars', are not easily identified in food or drink labels. Certain manufactured foods and drinks with 'safe' names, such as dried fruit and fruit juice, still contain free sugars and can be confusing. Guidance states that daily consumption of free sugars should be less than 10% of total energy intake (no more than 5% in the UK). However, it is found that both tooth decay and obesity are associated with consumption of free sugars in large quantities and at inappropriate times.

Removal of Water-Soluble Extractives Improves the Enzymatic Digestibility of Steam-Pretreated Softwood Barks.

Science.gov (United States)

Frankó, Balázs; Carlqvist, Karin; Galbe, Mats; Lidén, Gunnar; Wallberg, Ola

2018-02-01

Softwood bark contains a large amounts of extractives-i.e., soluble lipophilic (such as resin acids) and hydrophilic components (phenolic compounds, stilbenes). The effects of the partial removal of water-soluble extractives before acid-catalyzed steam pretreatment on enzymatic digestibility were assessed for two softwood barks-Norway spruce and Scots pine. A simple hot water extraction step removed more than half of the water-soluble extractives from the barks, which improved the enzymatic digestibility of both steam-pretreated materials. This effect was more pronounced for the spruce than the pine bark, as evidenced by the 30 and 11% glucose yield improvement, respectively, in the enzymatic digestibility. Furthermore, analysis of the chemical composition showed that the acid-insoluble lignin content of the pretreated materials decreased when water-soluble extractives were removed prior to steam pretreatment. This can be explained by a decreased formation of water-insoluble "pseudo-lignin" from water-soluble bark phenolics during the acid-catalyzed pretreatment, which otherwise results in distorted lignin analysis and may also contribute to the impaired enzymatic digestibility of the barks. Thus, this study advocates the removal of extractives as the first step in the processing of bark or bark-rich materials in a sugar platform biorefinery.

The chemosensitivity of labellar sugar receptor in female Phormia regina is paralleled with ovary maturation: Effects of serotonin.

Science.gov (United States)

Solari, Paolo; Stoffolano, John G; De Rose, Francescaelena; Barbarossa, Iole Tomassini; Liscia, Anna

2015-11-01

Oogenesis in most adult insects is a nutrient-dependent process involving ingestion of both proteins and carbohydrates that ultimately depends on peripheral input from chemoreceptors. The main goal of this study was to characterize, in the female blowfly Phormia regina, the responsive changes of the labellar chemoreceptors to carbohydrates and proteins in relation to four different stages along the ovarian cycle: (1) immature ovaries, (2) mid-mature ovaries, (3) mature ovaries and ready for egg-laying and (4) post egg-laying ovaries. Then, the possible effects exerted by exogenous serotonin on the chemoreceptor sensitivity profiles were investigated. Our results show that ovary length, width and contraction rate progressively increase from stage 1 to 3, when all these parameters reach their maximum values, before declining in the next stage 4. The sensitivity of the labellar "sugar" chemoreceptors to both sucrose and proteins varies during the ovarian maturation stages, reaching a minimum for sucrose in stage 3, while that to proteins begins. Exogenous 5-HT supply specifically increases the chemoreceptor sensitivity to sugar at the stages 3 and 4, while it does not affect that to proteins. In conclusion, our results provide evidence that in female blowflies the cyclic variations in the sensitivity of the labellar chemosensilla to sugars and proteins are time-related to ovarian development and that during the stages 3 and 4 the responsiveness of the sugar cell to sucrose is under serotonergic control. Copyright © 2015 Elsevier Ltd. All rights reserved.

Nutrient Intake and Digestibility of Cynomolgus Monkey (Macaca fascicularis Fed with High Soluble Carbohydrate Diet: A Preliminary Study

Directory of Open Access Journals (Sweden)

DEWI APRI ASTUTI

2009-12-01

Full Text Available High carbohydrate as obese diet is not yet available commercially for monkeys. Therefore, this preliminary study was to carry out nutrient intake and digestibility of cynomolgus monkeys (Macaca fascicularis fed with high soluble carbohydrate diet compared to monkey chow. Five adult female macaques (average body weight 2.67 kg were made to consume freshly diet. Commercial monkey chows (contains 3500 cal/g energy and 35% starch were fed to three adult females (average body weight 3.62 kg. Nutrient intakes and digestibility parameters were measured using modified metabolic cages. Result showed that average of protein, fat, starch, and energy intakes in treatment diet were higher than control diet (T-test. Fat intake in the treatment diet was three times higher, while starch and energy intakes were almost two times higher than monkey chow. Digestibility percentage of all nutrients were the same in both diets except for the protein. The study concludes that the freshly prepared high sugar diet was palatable and digestible for the cynomolgus monkeys. Further studies are in progress to develop obese diet high in energy content based on fat and source of starch treatments.

Sugar-Sweetened Beverages Are the Main Sources of Added Sugar Intake in the Mexican Population.

Science.gov (United States)

Sánchez-Pimienta, Tania G; Batis, Carolina; Lutter, Chessa K; Rivera, Juan A

2016-09-01

Sugar intake has been associated with an increased prevalence of obesity, other noncommunicable diseases, and dental caries. The WHO recommends that free sugars should be ENSANUT (National Health and Nutrition Survey) 2012], which represents 3 geographic regions and urban and rural areas. Dietary information was obtained by administering a 24-h recall questionnaire to 10,096 participants. Total sugar intake was estimated by using the National Institute of Public Health (INSP) food-composition table and an established method to estimate added sugars. The mean intakes of total, intrinsic, and added sugars were 365, 127, and 238 kcal/d, respectively. Added sugars contributed 13% of TEI. Sugar-sweetened beverages (SSBs) were the main source of sugars, contributing 69% of added sugars. Food products high in saturated fat and/or added sugar (HSFAS) were the second main sources of added sugars, contributing 25% of added sugars. The average intake of added sugars in the Mexican diet is higher than WHO recommendations, which may partly explain the high prevalence of obesity and diabetes in Mexico. Because SSBs and HSFAS contribute >94% of total added sugars, strategies to reduce their intake should be strengthened. This includes stronger food labels to warn the consumer about the content of added sugars in foods and beverages. © 2016 American Society for Nutrition.

Soluble polymorphic bank vole prion proteins induced by co-expression of quiescin sulfhydryl oxidase in E. coli and their aggregation behaviors.

Science.gov (United States)

Abskharon, Romany; Dang, Johnny; Elfarash, Ameer; Wang, Zerui; Shen, Pingping; Zou, Lewis S; Hassan, Sedky; Wang, Fei; Fujioka, Hisashi; Steyaert, Jan; Mulaj, Mentor; Surewicz, Witold K; Castilla, Joaquín; Wohlkonig, Alexandre; Zou, Wen-Quan

2017-10-04

The infectious prion protein (PrP Sc or prion) is derived from its cellular form (PrP C ) through a conformational transition in animal and human prion diseases. Studies have shown that the interspecies conversion of PrP C to PrP Sc is largely swayed by species barriers, which is mainly deciphered by the sequence and conformation of the proteins among species. However, the bank vole PrP C (BVPrP) is highly susceptible to PrP Sc from different species. Transgenic mice expressing BVPrP with the polymorphic isoleucine (109I) but methionine (109M) at residue 109 spontaneously develop prion disease. To explore the mechanism underlying the unique susceptibility and convertibility, we generated soluble BVPrP by co-expression of BVPrP with Quiescin sulfhydryl oxidase (QSOX) in Escherichia coli. Interestingly, rBVPrP-109M and rBVPrP-109I exhibited distinct seeded aggregation pathways and aggregate morphologies upon seeding of mouse recombinant PrP fibrils, as monitored by thioflavin T fluorescence and electron microscopy. Moreover, they displayed different aggregation behaviors induced by seeding of hamster and mouse prion strains under real-time quaking-induced conversion. Our results suggest that QSOX facilitates the formation of soluble prion protein and provide further evidence that the polymorphism at residue 109 of QSOX-induced BVPrP may be a determinant in mediating its distinct convertibility and susceptibility.

Analysis of biochemical characteristics of maize anthers from male fertilizable plant and male sterile plant induced by space flight

International Nuclear Information System (INIS)

Zhang Lingbi; Rong Tingzhao; Zhang Caibo; Sichuan Agricultural Univ., Ya'an; Zhu Yingguo; Cao Moju; Sichuan Agricultural Univ., Ya'an; Wuhan Univ., Wuhan

2007-01-01

The contents of soluble sugar, starch, protein and free praline in anthers were analyzed and compared between male fertilizable and male sterile induced by space flight at different developmental stages. The results showed that all contents were lower in male sterile anthers than in fertilisable ones. With anther developing, the contents of soluble starch, protein and free proline increased obviously in male fertilizable anthers, but not in sterile ones. (authors)

75 FR 60715 - Domestic Sugar Program-FY 2010 and FY 2011 Cane Sugar and Beet Sugar Marketing Allotments and...

Science.gov (United States)

2010-10-01

... marketing allotment and the associated production history will be transferred from MDFC to WSG, effective... Cane Sugar and Beet Sugar Marketing Allotments and Company Allocations AGENCY: Commodity Credit... publish the modifications to the fiscal year 2010 (FY 2010) State sugar marketing allotments and company...

Organic compounds in hot-water-soluble fractions from water repellent soils

Science.gov (United States)

Atanassova, Irena; Doerr, Stefan

2014-05-01

Water repellency (WR) is a soil property providing hydrophobic protection and preventing rapid microbial decomposition of organic matter entering the soil with litter or plant residues. Global warming can cause changes in WR, thus influencing water storage and plant productivity. Here we assess two different approaches for analysis of organic compounds composition in hot water extracts from accelerated solvent extraction (ASE) of water repellent soils. Extracts were lyophilized, fractionated on SiO2 (sand) and SPE cartridge, and measured by GC/MS. Dominant compounds were aromatic acids, short chain dicarboxylic acids (C4-C9), sugars, short chain fatty acids (C8-C18), and esters of stearic and palmitic acids. Polar compounds (mainly sugars) were adsorbed on applying SPE clean-up procedure, while esters were highly abundant. In addition to the removal of polar compounds, hydrophobic esters and hydrocarbons (alkanes and alkenes particle wettability and C dynamics in soils. Key words: soil water repellency, hot water soluble carbon (HWSC), GC/MS, hydrophobic compounds

21 CFR 173.320 - Chemicals for controlling microorganisms in cane-sugar and beet-sugar mills.

Science.gov (United States)

2010-04-01

...-sugar and beet-sugar mills. 173.320 Section 173.320 Food and Drugs FOOD AND DRUG ADMINISTRATION... controlling microorganisms in cane-sugar and beet-sugar mills. Agents for controlling microorganisms in cane-sugar and beet-sugar mills may be safely used in accordance with the following conditions: (a) They are...

pH-sensitive polymeric nanoparticles to improve oral bioavailability of peptide/protein drugs and poorly water-soluble drugs.

Science.gov (United States)

Wang, Xue-Qing; Zhang, Qiang

2012-10-01

pH-sensitive polymeric nanoparticles are promising for oral drug delivery, especially for peptide/protein drugs and poorly water-soluble medicines. This review describes current status of pH-sensitive polymeric nanoparticles for oral drug delivery and introduces the mechanisms of drug release from them as well as possible reasons for absorption improvement, with emphasis on our contribution to this field. pH-sensitive polymeric nanoparticles are prepared mainly with polyanions, polycations, their mixtures or cross-linked polymers. The mechanisms of drug release are the result of carriers' dissolution, swelling or both of them at specific pH. The possible reasons for improvement of oral bioavailability include the following: improve drug stability, enhance mucoadhesion, prolong resident time in GI tract, ameliorate intestinal permeability and increase saturation solubility and dissolution rate for poorly water-soluble drugs. As for the advantages of pH-sensitive nanoparticles over conventional nanoparticles, we conclude that (1) most carriers used are enteric-coating materials and their safety has been approved. (2) The rapid dissolution or swelling of carriers at specific pH results in quick drug release and high drug concentration gradient, which is helpful for absorption. (3) At the specific pH carriers dissolve or swell, and the bioadhesion of carriers to mucosa becomes high because nanoparticles turn from solid to gel, which can facilitate drug absorption. Copyright © 2012 Elsevier B.V. All rights reserved.

Aluminum solubility and mobility in relation to organic carbon in surface soils affected by six tree species of the northeastern United States

NARCIS (Netherlands)

Dijkstra, F.A.; Fitzhugh, R.D.

2003-01-01

We compared Al solubility and mobility in surface soils among six tree species (sugar maple [Acer saccharum], white ash [Fraxinus americana], red maple [Acer rubrum, L.], American beech [Fagus grandifolia, Ehrh.], red oak [Quercus rubra, L.], and hemlock [Tsuga canadensis, Carr.]) in a mixed

Quantificação de açúcares solúveis totais, açúcares redutores e amido nos grãos verdes de cultivares de milho na colheita Quantification of total soluble sugars, reducing sugars and starch in immature kernels of corn cultivars

Directory of Open Access Journals (Sweden)

Fernanda Fátima Caniato

2007-12-01

Full Text Available O objetivo deste trabalho foi quantificar os teores de açúcares solúveis totais (AST, açúcares redutores (AR e amido em nove cultivares de milho colhidos no estádio verde, uma vez que estes influenciam diretamente o sabor, o aroma e a aceitação do consumidor. O experimento foi conduzido em Ponte Nova, MG, no ano agrícola 2002/2003. Para a determinação das características, foi retirada uma amostra de três espigas de cada unidade experimental. O ponto de colheita foi identificado por método empírico, quando o estilo-estigma se desprendia da espiga com facilidade. As cultivares apresentaram umidades diferentes no momento da colheita, variando de 56 a 64%. Não foi detectada diferença entre as cultivares quanto aos teores de AR, porém, P3232 e AG4051 apresentaram boa relação entre AR e AST, destacando-se para esta finalidade.Total soluble sugars (TSS, reducing sugars (RS, moisture and starch directly influence flavor and consumption. In this work, TSS, RS and starch were determined at harvest in young kernels of nine corn cultivars. The experiment was carried out in Ponte Nova, MG, Brazil. To determine chemical characters of young kernels a sample was obtained of three cobs from each experimental plot. Corn ears were harvested using an empirical method, when the silk was loose from the ear. Moisture varied among samples of different cultivars at harvest. Moisture content ranged from 56 to 64%. There were no differences among cultivars regarding RS. However, cultivars P3232 and AG4051 showed good RS to TSS ratio, being adequate for fresh consumption.

Biological variation and reference intervals for circulating osteopontin, osteoprotegerin, total soluble receptor activator of nuclear factor kappa B ligand and high-sensitivity C-reactive protein

DEFF Research Database (Denmark)

Sennels, H P; Jacobsen, Søren; Jensen, T

2007-01-01

Objective. Monitoring inflammatory diseases and osteoclastogenesis with osteopontin (OPN), osteoprotegerin (OPG), total soluble receptor activator of nuclear factor kappa B ligand (total sRANKL) and high-sensitivity C-reactive protein (hsCRP) has recently attracted increased interest. The purpose...

Damage to the microbial cell membrane during pyrolytic sugar utilization and strategies for increasing resistance.

Science.gov (United States)

Jin, Tao; Rover, Marjorie R; Petersen, Elspeth M; Chi, Zhanyou; Smith, Ryan G; Brown, Robert C; Wen, Zhiyou; Jarboe, Laura R

2017-09-01

Lignocellulosic biomass is an appealing feedstock for the production of biorenewable fuels and chemicals, and thermochemical processing is a promising method for depolymerizing it into sugars. However, trace compounds in this pyrolytic sugar syrup are inhibitory to microbial biocatalysts. This study demonstrates that hydrophobic inhibitors damage the cell membrane of ethanologenic Escherichia coli KO11+lgk. Adaptive evolution was employed to identify design strategies for improving pyrolytic sugar tolerance and utilization. Characterization of the resulting evolved strain indicates that increased resistance to the membrane-damaging effects of the pyrolytic sugars can be attributed to a glutamine to leucine mutation at position 29 of carbon storage regulator CsrA. This single amino acid change is sufficient for decreasing EPS protein production and increasing membrane integrity when exposed to pyrolytic sugars.

Improved sugar beet pectin-stabilized emulsions through complexation with sodium caseinate.

Science.gov (United States)

Li, Xiangyang; Fang, Yapeng; Phillips, Glyn O; Al-Assaf, Saphwan

2013-02-13

The study investigates the complexes formed between sodium caseinate (SC) and sugar beet pectin (SBP) and to harness them to stabilize SBP emulsions. We find that both hydrophobic and electrostatic interactions are involved in the complexation. In SC/SBP mixed solution, soluble SC/SBP complexes first form on acidification and then aggregate into insoluble complexes, which disassociate into soluble polymers upon further decreasing pH. The critical pH's for the formation of soluble and insoluble complexes and disappearance of insoluble complexes are designated as pH(c), pH(φ), and pH(d), respectively. These critical pH values define four regions in the phase diagram of complexation, and SC/SBP emulsions were prepared in these regions. The results show that the stability of SBP-stabilized emulsion is greatly improved at low SC/SBP ratios and acidic pH's. This enhancement can be attributed to an increase in the amount of adsorbed SBP as a result of cooperative adsorption to sodium caseinate. Using a low ratio of SC/SBP ensured that all caseinate molecules are completely covered by adsorbed SBP chains, which eliminates possible instability induced by thermal aggregation of caseinate molecules resulting from stress acceleration at elevated temperatures. A mechanistic model for the behavior is proposed.

A physiological and genetic approach to the improvement of tomato (Lycopersicon esculentum Mill.) fruit soluble solids

International Nuclear Information System (INIS)

Damon, S.E.

1989-01-01

Physiological processes and the genetic basis determining soluble solids content (SSC) of processing tomato fruit were addressed. Analysis of [ 3 H]-(fructosyl)-sucrose translocation in tomato indicates that phloem unloading in the fruit occurs, at least in part, to the apoplast. Apoplastic sucrose, glucose and fructose concentrations were estimated as 1 to 7, 12 to 49 and 8 to 63 millimolar, respectively in tomato fruit pericarp. Short-term uptake of [ 14 C]sucrose, -glucose and -fructose in tomato pericarp discs showes first order kinetics over the physiologically relevant concentration range. The uptake of [ 14 C]-(glycosyl)-1'fluorosucrose was identical to the rate of [ 14 C] sucrose uptake suggesting sucrose may be taken up directly without prior extracellular hydrolysis. Short-term uptake of all three sugars was insensitive to 10 micromolar carbonyl cyanide m-chlorophenylhydrazone and to 10 micromolar p-chloromercuribenzene sulfonic acid. However, long-term accumulation of glucose was sensitive to carbonyl cyanide m-chlorophenylhydrazone. Sugar uptake across the plasmamembrane does not appear to be energy dependent, suggesting that sugar accumulation in the tomato is driven by subsequent intracellular metabolism and/or active uptake at the tonoplast. Fourteen genomic DNA probes and ten restriction endonucleases were used to identify restriction fragment length polymorphisms (RFLPs) useful in the linkage analysis of quantitative trait loci controlling the expression of SSC in a segregating F 2 population from a cross between L. esculentum (UC204B) and L. cheesmanii f. minor, a wild species with high fruit soluble solids. RFLPs were detected between the DNAs of the two tomato species with all 14 probes

One-step enzymatic synthesis of nucleosides from low water-soluble purine bases in non-conventional media.

Science.gov (United States)

Fernández-Lucas, Jesús; Fresco-Taboada, Alba; de la Mata, Isabel; Arroyo, Miguel

2012-07-01

The effect of several water-miscible cosolvents on activity and stability of soluble and immobilized 2'-deoxyribosyltransferase from Lactobacillus reuteri on Sepabeads® has been studied in order to establish optimal conditions for enzymatic synthesis of nucleosides using purine bases with low solubility in aqueous buffer. As a rule of thumb, there was a general reduction of soluble enzyme activity when cosolvent content was gradually increased in reaction medium. In contrast, immobilized enzyme activity was enhanced 1.2-1.4-fold at 20% of methanol, ethanol, 2-propanol, diethylene glycol, and acetone; and at 10% and 30% acetonitrile. Likewise, highest increased activity (1.8-fold) was also obtained in presence of 20% acetonitrile. Immobilized enzyme was successfully used in the synthesis of 2'-deoxyxanthosine and 2'-deoxyguanosine using 2'-deoxyuridine as sugar donor and the corresponding poor water-soluble base in the presence of 30% of methanol, ethanol, 2-propanol, ethylene glycol, acetonitrile, and DMSO, giving high nucleoside yields at 4h. Copyright © 2011 Elsevier Ltd. All rights reserved.